Judul : Jodoh dari Negeri Seberang; Catatan Cinta Pernikahan Beda Bangsa
Penulis : Rahmadiyanti Rusdi dkk
Penerbit : Lingkar Pena Publishing House
Tahun : 2009
Genre : Pernikahan
Tebal : 240 Halaman
ISBN : 978-979-1367-83-7
By Wisata Buku.com:
Untuk membuka lembaran hidup baru, jalan berliku harus dilalui. Dengan calon pasangan, ada hal-hal yang perlu didiskusikan: pandangan hidup, cita-cita, rencana masa depan, konsep keluarga, sampai pengelolaan ekonomi dan pengasuhan anak, tentu saja berikut tempat tinggal. Dengan keluarga, harus pula presentasi bersama karena akan sukar menempuh biduk lanjutan ini tanpa restu khususnya dari orangtua kedua belah pihak.
Orangtua dahulu (dalam bahasa Sunda) mengatakan bahwa jodoh itu “jorok”. Terjemahan bebasnya, jodoh adalah misteri. Kita tak bisa menggunakan rancangan detail bagaikan arsitek, tak bisa menggunakan pola USP seperti hendak meluncurkan produk baru, tak bisa juga menggunakan analisis statistik regresi atau berpandu pada variabel terikat dan tidak terikat. Karena demikian abstraknya, proses pertemuan dengan sang jodoh mendebarkan sekaligus mengusik emosi. Simak saja tuturan para kontributor dalam buku ini.
Berjumpa belahan hati dari benua yang berjauhan, tak bisa ditampik bila memang sudah garis Yang Maha Kuasa. Istri-istri berkumpul di buku ini menuangkan suka-dukanya, terutama urusan birokrasi karena begitu melekat cap berkelimpahan materi pada orang asing sehingga dianggap “layak” dipersukar dan dikenai tarif yang menggelembung dari biasanya. Padahal palu belum final. Di negeri yang hendak dijadikan kediaman kelak, masih menanti aneka pernik adaptasi. Bukan hanya dengan budaya sekitar, pemahaman keluarga suami terhadap muslim dan sejumlah perbedaan personal yang perlu dijembatani mengingat rata-rata kontributor saling mengenal melalui dunia maya pun harus dilalui seserius mungkin.
Dibandingkan Nikah Sama Bule, yang penyusun dan penerbitnya sama, saya lebih menyukai Jodoh dari Negeri Seberang. Dari segi judul, cenderung berkonotasi “dewasa” karena tidak terlalu gaul. Klop dengan foto di kulit depannya yang indah, mengedepankan kecantikan cinta kala diikat di pelaminan dan menyatakan jelas bahwa kisah-kisah ini benar adanya. Masih dengan sisipan tips dan foto para kontributor beserta keluarga masing-masing, Jodoh dari Negeri Seberang enak dibaca lagi mengandung sejumlah pengetahuan yang dibutuhkan bagi Anda yang bersiap meniti dunia rumahtangga di rantau.
Ingin tahu di mana itu Comoro? Bagaimana mengharmoniskan diri dengan pasangan dari Turki? Bagaimana merenda hari-hari di Tokyo? Apakah pria asal Colombia memang semesra tokoh-tokoh sentral di telenovela? Cari tahu di buku ini
By Haryadi
Sesuai judulnya, buku yang ditulis oleh 11 penulis ini (semuanya wanita), mengisahkan betapa unik dan nano-nanonya melakukan pernikahan dengan warga negara asing. Jangankan dengan warga negara asing yang tentu serba ’asing’ (bahkan Mbak Hani –salah satu penulis buku ini, menyebutnya sebagai Alien, hehe), tak sedikit orang yang berpendapat bahwa menyatukan dua manusia dalam ikatan pernikahan (satu suku, satu bangsa dan negara sekalipun) itu susah. Bukan sebuah hal yang mudah bagi penulis-penulis buku ini, terlebih ketika mereka harus menyatukan perbedaan kultur yang serba baru. Hampir semua penulis di buku ini mengakui hal itu. Bahkan, tak sedikit yang mengaku sempat terkena culture shock
Tulisan pertama yang aku baca berjudul ”Pinangan dari Le Perisien” yang dibuat oleh Mbak Ita . Jujur aja, selain Mbak Dee yang duduk dibangku penyusun, cuma Mbak Ita yang aku kenal. Melalui blognya, cukup banyak tulisan Mbak Ita yang berkisah mengenai pernikahannya dengan Mas Pat. Untungnya, dibuku ini, Mbak Ita menulis hal baru. Asyik sekali mengetahui secara detail ketika Mas Pat melamar mbak Ita diantara rintikan hujan. Aih romantisnya. 🙂
Selanjutnya aku kembali baca dari awal. Tulisan Mbak Suci Al-Sadiq yang berjudul ”From India With Love” bikin aku merinding. Bukan karena serem, tapi karena betapa teguh dan mantapnya keyakinan Mbak Suci untuk memutuskan menikah dengan lelaki yang hanya ia kenal lewat internet.
Aku tidak mau menyebutkan satu persatu kisah penulis yang ada di buku ini. Yang pasti, aku bisa menikmati semua kata demi kata yang sudah dituturkan. Beberapa cerita cenderung sama (misalnya ketika mendapat pertentangan dari orang tua) tapi masing-masing bisa diceritakan dengan cara yang berbeda dan khas. Lebih asyik lagi ketika mengetahui hal-hal unik yang biasa dilakukan suami mereka (berhubungan dengan kebiasaan negara asal suami mereka). Mungkin akan lebih asyik kalo ada satu penulis pria yang menikah dengan wanita WNA. *kaya si Oji itu tuh, hehe* banyak juga, kan, pria Indonesia yang menikah dengan WNA.
Udah lama aku gak beli buku-bukunya LPPH. Kaget, karena secara kualitas fisik, rada menurun dari buku-buku sebelumnya *zaman-zaman Matahari Tak Pernah Sendiri* hmm, mungkin untuk menekan harga kali, ya? asyiknya, masing-masing cerita disertai foto dan keterangan foto (ada satu foto yang tidak diberi keterangan, aku lupa halaman berapa). Dan sayangnya, lumayan banyak kesalahan cetak dibuku ini. Misalnya saja di halaman 136, kata ”kesedihanku” ditulis ”kesedihankuku”. Atau di halaman 115 di paragraf 4. Semoga kedepan hal kecil yang cukup mengganggu ini bisa diperbaiki
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In order to grow bacteria Escherichia coli, we need to prepare a nutrient liquid media called LB or Luria-Bertani. Here are the steps in preparing it:
1. Weigh:
1g Tripton,
0,5 g yeast
1g NaCl
2. Put all in a 200 ml beaker, add aquadest to 100 mL
3. Mix and heat on a hotplate with magnetic stirrer until you get clean yellow solution
3. move the mixture into a 250 ml erlenmeyer, cover it with aluminum foil
4. Sterilize LB mixture in Erlenmeyer with an autoclave in 121C for 20 minutes
5. keep in 4C until you need it
good luck
A designed primer can stick to RNA template
Picture source: worthington-biochem.com
A designed primer can stick to DNA template too
Primer Specific Gene
picture source: bio.davidson.edu
Primers specific gene are needed if we want to characterize sequence of a certain enzyme encoding gene by generating copy-DNA from the RNA template or simply from DNA templates if available. For example if you want to characterize gene A from plant B, you can utilize submitted complete gene code of gene A from other plants. We can take benefits from conserve area of the gene of different plants as the basic for designing primers. If you begin from the information about the enzyme expressed by the gene that means you will have to isolate the RNA and use it as a template to generate cDNA that represents a the RNA encoding gene. Here are the basic steps to do it:
1. Visit gene-bank website and type your search in protein are, type the name of enzyme whose gene you want to characterize.
2. you may find lists of similar enzyme but from different plant species. find the FASTA file type of set of amino acids from every enzyme listed there.
3. using clustalW software, align those FASTA codes of your enzyme coming from several plants. so in the input you will place several sets of FASTA codes obtained from different plant species.
4. find conserve area of the enzyme (RNA) displayed by an array of stars.
5. amino acid codes array that at least have 7 amino acids (or 3×7 base pair because 1 amino acid consist of 3 bases in a codon) can be used to design a primer.
picture source: topnews.in
ADVANCE ENGLISH:
Lesson Plan 1:
The warm-up: I greet my students. “Hi, everyone. How are you today?” Did you have a good weekend?” Such casual exchange makes for a good warm-up, and students react well to it.
The Review: I ask the students:” Does anyone have any questions on the last lesson? “I would answer their questions, and then go over the outline for the day’s lesson.
The presentation: I start with an introduction to the topic, then explain the lesson, encouraging students to ask questions by suggesting,”Does anyone have a question? Please ask. I am here to help you.” I put forward the topic for discussion and ask students to split into groups, before I engage them in conversation, writing their and my ideas on the board.
The pronunciation: I have my students repeat aloud words and sentences after me.
The closing: I encourage students to ask questions one last time, before ending with a song, whose vocabulary I explain. I assign my students five sentences to write in English. During the next lesson, I would have each student write his sentences on the board. Other students and I would take part in correcting those sentences, thus giving all the students the opportunity to recognize mistakes, as well as their corrections. This would enhance learning.
Lesson Plan 2:
The Warm-up: I greet my students. “How are you doing today? Everybody’s looking good.” This ensures a warm and positive response.
The Review: I answer questions and go over the lesson structure with the students.
The Presentation: I start with a question, or two, related to the conversation top and to students lives, as well e.g. if we’re talking about traffic congestion, I ask, “Have you experienced any delays on your way to work in the morning because of traffic jams?”
The pronunciation: I play an English lesson CD and have the students repeat after the speaker in unison.
The closing: I divide the students into groups, draw pictures of different items on the board and ask them to guess the English word for each one, while I keep score. For beginning – level students I would assign them a paragraph to write making top suggestions and helping then form a few sentences before they write it.
Lesson plan 3
The warm-up: I start a chat about yesterday’s activities. This would give a good warm-up, as well as an opportunity to teach a few new words.
The review: I encourage students to ask a question on the last lesson, as well as anything on their mind, which is related to the English language and take in their questions.
The Presentations: Occasionally, I would have a student prepare a presentation. They would take the floor and explain their researched work. I would help that student take questions.
The pronunciation: I have the students repeat greetings and other important, everyday English after me.
The closing: I would use an illustrated book and ask students for the meaning of the objects within its page. For more advanced students I would assign them an essay to write, explaining how to structure it into an introduction, body paragraphs and conclusion. I would take this essay home with me to correct to give back to students with corrections and remark.
Lesson Plan 4:
The warm –up: I would mention the topic for the day’s lesson, whether it is based on a book, or chosen by me for conversation, and ask students what they know about it.
The review: I would mention the topic for the day’s lesson whether it is based on a book or chosen by me for conversation and ask students what they know about it.
The presentation: I would break down the lesson into several points and address each one separately, before summing it up and taking questions.
The pronunciation: I ask students if they have a difficult pronouncing any words, or character, and I help them pronounce them.
The closing: I would close with a game, asking each student to describe his classmate. For home assignment I would ask students to listen to a song of their choice and rewrite its lyrics. This would be a good, stimulating exercise, which would enrich their vocabulary and writing skills. I would encourage them to use a good dictionary.
Lesson plan 5
The warm up: I’d talk about the weather, using the opportunity to teach students weather-related vocabulary.
The review: I would give students the opportunity to choose the conversation topic from several choices.
The presentation: Having prepared the day’s lesson beforehand, I follow the book’s lesson plan alternating assigning students reading material and playing CD, while asking students questions and answering theirs.
The pronunciation: I have the students pronounce words five times after me to make sure their learning well.
The closing: I close by asking students to say or write his favorite quote one extracurricular activity, which I have actually done while teaching is to go out with students to a café, or visit a touristic or archaeological site, while speaking only English.
Lesson plan 1
The Warm-up: I start a chat about today’s news on newspaper and what they feel about it. This would bring positive respond, as well as an opportunity to teach a few new words about forecast.
The Review: I encourage students to ask a question on the last lesson, as well as anything on their mind, which is related to the English language and take in their questions.
The Presentations: Having prepared the day’s lesson beforehand, I follow the book’s lesson plan alternating assigning students reading material and playing recorded tape, while asking students questions and answering theirs.
The Pronunciation: I have the students repeat important reports and other important everyday English after me.
The Closing: I would use an illustrated diagram and ask students for the information we can get within its page. For more advanced students I would assign them an essay to write, explaining how to structure it into an introduction, body paragraphs and conclusion. I would take this essay home with me to correct to give back to students with corrections and remark.
Lesson Plan 2
The Warm-up: I’d talk about hobbies and other daily activities at home. This ensures a warm and positive response.
The Review: I answer questions and go over the lesson structure with the students.
The Presentation: I start with a question, or two, related to the conversation top and to students lives, as well e.g. if we’re talking about household, I ask, “Have you experienced any electrical shock when you do chorus and how to avoid that?”
The Pronunciation: I play an English lesson CD and have the students repeat after the speaker in unison.
The Closing: I divide the students into groups, draw pictures of different items on the board and ask them to guess the English word for each one, while I keep score. For beginning – level students I would assign them a paragraph to write making top suggestions and helping then form a few sentences before they write it.
Lesson Plan 3
The Warm-up: I greet my students. “Hello everyone. How is everything going?” Do you have a good plan for this weekend?” Such casual exchange makes for a good warm-up, and students react well to it.
The Review: I ask the students:” Does anyone have any questions on the last lesson? “I would answer their questions, and then go over the outline for the day’s lesson that I pick from their textbook.
The Presentation: I start with an introduction to the topic, then explain the lesson, encouraging students to ask questions by suggesting, ”Does anyone have a question? Please raise your hand. I am here if you need me to help you.” I put forward the topic for discussion and ask students to split into groups, before I engage them in conversation, writing their and my ideas on the board.
The Pronunciation: I have my students repeat aloud words and sentences after me.
The Closing: I encourage students to ask questions one last time, before ending with a song, whose vocabulary I explain. I assign my students five sentences to write in English. During the next lesson, I would have each student write his sentences on the board. Other students and I would take part in correcting those sentences, thus giving all the students the opportunity to recognize mistakes, as well as their corrections. This would enhance learning.
Lesson plan 4
The Warm –up: I would mention the topic of the day, for example about transportation. It is based on a book, or chosen by me for conversation, and ask students what they know about it.
The Review: I would give students the opportunity to choose the conversation topic from several choices.
The Presentation: I would break down the lesson into several points and address each one separately, before summing it up and taking questions.
The Pronunciation: I ask students to write 2 or 3 words or characters that they have difficulties to pronounce and I help them pronounce them.
The Closing: I would close with a bingo game, asking each student to guess silent-word acts performed by other student. For home assignment I would ask students to listen to a song of their choice and rewrite its lyrics. This would be a good, stimulating exercise, which would enrich their vocabulary and writing skills. I would encourage them to use a good dictionary.
Lesson Plan 5
The warm-up: I greet my students. “How are you today? You look all happy. What makes you so?” This ensures a warm and positive response.
The Review: I review homework and go over the lesson structure with the students.
The Presentation: I start with a question, or two, related to the conversation top and to students lives, as well e.g. if we’re talking about traffic congestion, I ask, “Have you experienced any delays on your way to work in the morning because of traffic jams?”
The Pronunciation: I play a short English song and have the students to repeat or sing together.
The Closing: The closing: I close by asking students to say or write his favorite dish along with recipe to make it one extracurricular activity, which I have actually done while teaching is to make a cooking demo, while speaking only English.
BUSINESS ENGLISH:
Lesson plan 1
The Audio-Lingual Method. Goal: collocation practice using words that begin with the letter B, practicing listening skills.
Review (5-10 min.). I will ask who remember some collocation we have already studied. Students will give examples. I will then explain that today’s lesson is collocation commonly used in business conversations.
Introduction (10 min). I will distribute a paper with one missing words then I will read each sentence and the class will repeat it. This will help them with the pronunciation of any unfamiliar words.
New material (25 min). Using an overhead, I will show them the words that they will need to fill in the blank. The students will work independently. When they are finished they will put the words in alphabetical order. When everyone is done I will read each sentence again and call on a different student to supply the answer and ask him to read the sentence. Then I will divide the class into pairs Student 1 will read one of the sentences we completed. Student 2 must respond to the statement in a way that makes sense. Example the customers have great brand loyalty for this product. Student 2 it must be the result of good ads. Then they switch roles. Continue switching roles until all the sentences have been answered. If a student gets stuck and can’t respond to the sentence, they get another try to respond to a different sentence.
Review/ Out of Class assignment (10 min). The assignment for the class will be to listen carefully to people talking in their place of business. They need to carry around a pen and a small pad of paper. Anytime they recognize someone using collocation at their place of work they need to write it down. They will bring their notepad to our next class and they will share what they heard. It for some reason they are not going to their job before our next class hey may read a short article on business and write down any collocation used in the article
I have just finished TESOL certification course with ATI. I took the thickest online program offered: Advanced TESOL Expert Online Certifications.
American TESOL Expert program is comprised of three courses totaling 200 hours. Successful graduates receive American TESOL Advanced certification, plus TESOL for Business and Children specialty certifications I received 3 e-books and 3 hard copy textbooks via DHL.
The course focuses on a variety of teaching approaches including the Rassias Method, the Total Physical Response Method, and the Communicative Approach. Using a combination of approaches is often a great way to ensure that your students are equally engaged. We have to submit assignments along with thesis to complete our program.
A week upon completion of this American TESOL program, I received TESOL certificate refering that I am qualified to teach any and all of the age groups, and be provided with job placement. I believe TESOL Expert Program is geared towards teaching children, adults, and business professionals.
This is the detail of program:
Program ID: ATIEP
Credentials Issued: TESOL Certificate + 2 Focus Certificates (Business & Children)
Length of Program: 200 hours
Delivery Method: Online Time Frame: At your own pace
I. Introduction
Enzymes are produced by all living cells as catalysts for specific chemical reactions. Not surprisingly enzymes are present in all foods at some time, and play an increasingly important role in food processing techniques. Enzymes, although not recognized as such, have played an essential part in some food processes, notably the making of cheese, leavened bread, wine and beer, for thousands of years (Dewdney, 1973).
II. Major Enzyme Applications in Food Industry
In food industry, enzyme has been used to produce and to increase the quality and the diversity of food. Some examples of products that use enzyme are cheese, yoghurt, bread syrup etc. Ancient traditional arts such as brewing, cheese making, meat tenderization with papaya leaves and condiment preparation (e.g., soy sauce and fish sauce) rely on proteolysis, albeit the methods were developed prior to our knowledge of enzymes. Early food processes involving proteolysis were normally the inadvertent consequence of endogenous or microbial enzyme activity in the foodstuff. Some major applications by types of enzymes are:
- Rennet
The use of rennet in cheese manufacture was among the earliest applications of exogenous enzymes in food processing, dating back to approximately 6000 B C. In 1994, the total production of cheese was 8000 metric tons against a total demand of 9000 metric tons. The projected demand by 2000 A D is around 30,000 metric tons. The use of rennet, as an exogenous enzyme, in cheese manufacture is perhaps the largest single application of enzymes in food processing. In recent years, proteinases have found additional applications in dairy technology, for example in acceleration of cheese ripening, modification of functional properties and preparation of dietic products (IDF, 1990).
Picture 1. Chymosin crystal structure (www.fst.rdg.ac.uk., 2002)
Animal rennet (bovine chymosin) is conventionally used as a milk-clotting agent in dairy industry for the manufacture of quality cheeses with good flavor and texture. Owing to an increase in demand for cheese production world wide – i.e. 4% per annum over the past 20 years, approximating 13.533 million tons (ref. 3) – coupled with reduced supply of calf rennet, has therefore led to a search for rennet substitutes, such as microbial rennet. At present, microbial rennet is used for one-third of all the cheese produced worldwide. Rennin acts on the milk protein in two stages, by enzymatic and by nonenzymatic action, resulting in coagulation of milk. In the enzymatic phase, the resultant milk becomes a gel due to the influence of calcium ions and the temperature used in the process (Bhoopathy, 1994). Many microorganisms are known to produce rennet-like proteinases which can substitute the calf rennet. Microorganisms like Rhizomucor pusillus, R. miehei, Endothia parasitica, Aspergillus oryzae, and Irpex lactis are used extensively for rennet production in cheese manufacture. Extensive research that has been carried out so far on rennet substitutes has been reviewed by several authors (Green, M. L., 1977 Fox, P. F.1993; Farkye, N. Y., 1995).
Different strains of species of Mucor are often used for the production of microbial rennets. Whereas best yields of the milk-clotting protease from Rhizomucor pusillus are obtained from semisolid cultures containing 50% wheat bran, R. miehei and Endothia parasitica are well suited for submerged cultivation. Using the former, good yields of milk-clotting protease may be obtained in a medium containing 4% potato starch, 3% soybean meal, and 10% barley. During growth, lipase is secreted together with the protease. Therefore, the lipase activity has to be destroyed by reducing the pH, before the preparation can be used as cheese rennet.
- Lactases
Lactose, the sugar found in milk and whey, and its corresponding hydrolase, lactase or b-galactosidase, have been extensively researched during the past decade (Mehaiya, 1987). This is because of the enzyme immobilization technique which has given new and interesting possibilities for the utilization of this sugar. Because of intestinal enzyme insufficiency, some individuals, and even a population, show lactose intolerance and difficulty in consuming milk and dairy products. Hence, low-lactose or lactose-free food aid programme is essential for lactose-intolerant people to prevent severe tissue dehydration, diarrhoea, and, at times, even death. Another advantage of lactase-treated milk is the increased sweetness of the resultant milk, thereby avoiding the requirement for addition of sugars in the manufacture of flavored milk drinks. Manufacturers of ice cream, yoghurt and frozen desserts use lactase to improve scoop and creaminess, sweetness, and digestibility, and to reduce sandiness due to crystallization of lactose in concentrated preparations. Cheese manufactured from hydrolyzed milk ripens more quickly than the cheese manufactured from normal milk.
Technologically, lactose crystallizes easily which sets limits to certain processes in the dairy industry, and the use of lactase to overcome this problem has not reached its fullest potential because of the associated high costs. Moreover, the main problem associated with discharging large quantities of cheese whey is that it pollutes the environment. But, the discharged whey could be exploited as an alternate cheap source of lactose for the production of lactic acid by fermentation. The whey permeate, which is a by-product in the manufacture of whey protein concentrates, by ultrafiltration could be fermented efficiently by Lactobacillus bulgaricus (Mehaiya, 1987)
Picture 3. Lactase crystal structure (Pitman, S. D., 2004)
Lactose can be obtained from various sources like plants, animal organs, bacteria, yeasts (intracellular enzyme), or molds. Some of these sources are used for commercial enzyme preparations. Lactase preparations from A. niger, A. oryzae, and Kluyveromyces lactis are considered safe because these sources already have a history of safe use and have been subjected to numerous safety tests. The most investigated E. coli lactase is not used in food processing because of its cost and toxicity problems.
The properties of the enzyme depend on its source. Temperature and pH optima differ from source to source and with the type of particular commercial preparation. Immobilization of the enzymes, method of immobilization, and type of carrier can also influence these optima values. In general, fungal lactase have pH optima in the acidic range 2.5–4.5, and yeast and bacterial lactases in the neutral region 6–7 and 6.5–7.5, respectively. The variation in pH optima of lactases makes them suitable for specific applications, for example fungal lactases are used for acid whey hydrolysis, while yeast and bacterial lactases are suitable for milk (pH 6.6) and sweet whey (pH 6.1) hydrolysis. Product inhibition, e.g. inhibition by galactose, is another property which also depends on the source of lactase. The enzyme from A. niger is more strongly inhibited by galactose than that from A. oryzae. This inhibition can be overcome by hydrolyzing lactose at low concentrations by using immobilized enzyme systems or by recovering the enzyme using ultrafiltration after batch hydrolysis. Lactases from Bacillus species are superior with respect to thermostability, pH operation range, product inhibition, and sensitivity against high-substrate concentration. Thermostable enzymes, able to retain their activity at 60°C or above for prolonged periods, have two distinct advantages viz. they give higher conversion rate or shorter residence time for a given conversion rate, and the process is less prone to microbial contamination due to higher operating temperature. Bacillus species have a pH optima of 6.8 and temperature optima of 65°C. Its high activity for skim milk and less inhibition by galactose has made it suitable for use as a production organism for lactase (Gekas, V. and Lopez-Levia, M., 1985).
The enzymatic hydrolysis of lactose can be achieved either by free enzymes, usually in batch fermentation process, or by immobilized enzymes or even by immobilized whole cells producing intracellular enzyme. Although numerous hydrolysis systems have been investigated, only few of them have been scaled up with success and even fewer have been applied at an industrial or semi-industrial level. Several acid hydrolysis systems have been developed to industry-scale level. Large-scale systems which use free enzyme process have been developed for processing of UHT-milk and processing of whey, using K. lactis lactase (Maxilact, Lactozyme).
Several commercial immobilized systems have been developed for commercial exploitation. Snamprogretti process of industrial-scale milk processing technology in Italy is one such working system. They make use of fibre-entrapped yeast lactase in a batch process, and the milk used is previously sterilized by UHT. For pilot plants, there are three other processes designed and developed to handle milk; (i) by Gist-Brocades, Rohm GmbH (Germany), and (ii) by Sumitomo, Japan. These are continuous processes with short residence times. Processing of whey UF-permeate is accomplished by the system developed by Corning Glass, Connecticut, Lehigh, Valio and Amerace corp. The process by Corning Glass is being applied at commercial scale in the bakers yeast production using hydrolyzed- whey (Gekas, V. and Lopez-Levia, M., 1985).
- Catalases
Catalase is an enzyme that can be produced from bovine livers or microbial sources. It is used to change hydrogen peroxide to become water and oxygen molecules. This enzyme can be used in a limited amount in cheese production. Catalase is the enzyme that breaks down hydrogen peroxide to water and molecular oxygen. Catalase effectively removes the residual hydrogen peroxide, ensuring that the fabric is peroxide-free and mainly used in food industry and also in egg processing with other enzymes. Catalase is a common enzyme found in nearly all living organisms which are exposed to oxygen, where it functions to catalyze the decomposition of hydrogen peroxide to water and oxygen (Chelikani P, Fita I, Loewen PC, 2004).
Picture 2. Catalase crystal structure in beef (green) (Mary Maj, 2009)
Glucose oxidase and catalase are often used together in selected foods for preservation. Superoxide dismutase is an antioxidant for foods and generates H2O2, but is more effective when catalase is present. Thermally induced generation of volatile sulphydryl groups is thought to be responsible for the cooked off-flavour in ultra high temperature (UHT) processed milk. Use of sulphydryl oxidase under aseptic conditions can eliminate this defect. The natural inhibitory mechanism in raw milk is due to the presence of low levels of lactoperoxidase (LP), which can be activated by the external addition of traces of H2O2 and thiocyanate. It has been reported that the potential of LP-system and its activation enhances the keeping quality of milk, (Muir, D. D., 1996)
- Lipases
A lipase is a water –soluble enzyme that catalyzes the hydrolysis of ester bonds in water-insoluble, lipid substrates (Svendsen, 2000). Lipases (triacylglycerol acylhydrolases) are produced by microorganism in individual or together with esterase. Microorganisms that produce lipases are Pseudomonas aeruginosa, Serratia marcescens, Staphylocococcus aureus dan Bacillus subtilis. Lipase is used as biocatalyst to produce free fatty acid, glycerol and various esters, part of glycerides and fat that is modified or esterified from cheap substrate i.e. palm oil. Those products are extensively used in pharmacy, chemical and food industry.
Picture 4. Lipase crystal structure (Berman, H. M., J. Westbrook, Z. Feng, G. Gilliland, T. N. Bhat, H. Weissig, I. N. Shindyalov, and P. E. Bourne, 2000)
Various animal or microbial lipases gave pronounced cheese flavor, low bitterness and strong rancidity, while lipases in combination with proteinases and/or peptidases give good cheese flavor with low levels of bitterness. In a more balanced approach to the acceleration of cheese ripening using mixtures of proteinases and peptidases, attenuated starter cells or cell-free extracts (CFE) are being favored (Wilkinson, M. G., 1995)
- Proteases
The proteolytic system of lactic acid bacteria is essential for their growth in milk, and contributes significantly to flavor development in fermented milk products. The proteolytic system is composed of proteinases which initially cleaves the milk protein to peptides; peptidases which cleave the peptides to small peptides and amino acids; and transport system responsible for cellular uptake of small peptides and amino acids. Lactic acid bacteria have a complex proteolytic system capable of converting milk casein to the free amino acids and peptides necessary for their growth. These proteinases include extracellular proteinases, endopeptidases, aminopeptidases, tripeptidases, and proline-specific peptidases, which are all serine proteases. Apart from lactic streptococcal proteinases, several other proteinases from nonlactostreptococcal origin have been reported. There are also serine type of proteinases, e.g. proteinases from Lactobacillus acidophillus, L. plantarum, L. delbrueckii sp. bulgaricus, L. lactis, and L. helveticus. Aminopeptidases are important for the development of flavor in fermented milk products, since they are capable of releasing single amino acid residues from oligopeptides formed by extracellular proteinase activity (Law, J., And Haandrikman, A., 1997)
Picture 5. Protease crystal structure (www.proteases.org, 2009)
- Amylases
They can be derived from bacteria and fungi. They play a major role in the food and beverages (baking), brewing, starch, sugar industries. Amylase is used to hydrolyze amilum into a product that is water soluble and has low molecular weight: glucose. This enzyme is used extensively in drink industry for example the production of High Fructose Syrup (HFS) or in textile industry. Amylases can be made from various microorganisms especially from Bacillus, Pseudomonas and Clostridium family. Potential bacteria that are recently used to produce amylases in industrial scale are Bacillus licheniformis and B.stearothermophillus. It is preferable to use B.stearothermophillus because it can produce enzyme that is thermo stable so that can reduce production cost.
Alpha amylases have significant effects on baked goods. If the content is low, this leads to low dextrin production and poor gas production. This in turn results in inferior quality bread with reduced size and poor crust color. To compensate for the deficiencies of the grain, it is necessary to add either sugar or alpha amylase. The addition of enzymes offers certain advantages over sugar. At a flour mill, it is possible to standardize the enzyme content of the flour so that a uniform commodity can be supplied. Furthermore, enzymes bring about a gradual formation of sugar, which matches the needs of the yeast. When the dough is placed in the oven, the steadily increasing temperature leads to an increase in the enzymes’ rate of reaction and more sugar is produced. Malt flour and malt extract can be used as enzyme supplements as malt is rich in alpha amylases. However, it is better to use a fungal alpha amylase.
Picture 6. Alpha-amylase crystal structure (www.polyenzyme.com, 2009)
The alpha-amylases degrade the damaged starch in wheat flour into small dextrins, thus allowing yeast to work continuously during dough fermentation, proofing and the early stage of baking. This results in improved bread volume and crumb texture. In addition, the small oligosaccharides and sugars such as glucose and maltose produced by these enzymes enhance the reactions for the browning of the crust and baked flavour. Cereal beta-amylases are perhaps best known in terms of the vital role they play in releasing easily fermentable sugars from cereal grain starch to fuel the production of alcohol by yeast in brewing. The extent to which they have been investigated is indeed largely due to their significance in this economically important industry. However, cereal beta-amylases are also, or could be, employed in many other aspects of the food industry and the analysis of starch, and they constitute valuable markers in cereal assessment and breeding studies (Ziegler, 1999).
References
Bhoopathy, R., 1994. Enzyme technology in food and health industries, Indian Food Ind., 13: 22–31
Chelikani, P., Fita, I., Loewen P. C., 2004. Diversity of structures and properties among catalases, Cell. Mol. Life Sci. 61 (2): 192-208.
Dewdney, P. A., 1973. Enzymes in food processing, Nutrition and Food Science Journal. 73 (4): 20–23.
Farkye, N. Y., 1995. Contribution of milk-clotting enzymes and plasmin to cheese ripening, Adv. Exp. Med. Biol., 367: 195–207
Gekas, V. and Lopez-Levia, M., 1985. Hydrolysis of lactose, a literature review, Process Biochem., 20: 2–12
Green, M. L., 1993. Review of the progress of dairy science: milk coagulants, J. Dairy Res., 1977, 44: 159–188.
Fox, P. F., 1998. Exogenous enzymes in dairy technology – a review, J. Food Biochem., 17: 173–175
IDF, Int. Dairy Fed. Bull., 1990. 247: 24–38.
Law, J. and Haandrikman, A., 1997. Proteolytic enzymes of lactic acid bacteria, Int. Dairy J., 7: 1–11.
Mehaia, M. A. and Cheryan, 1987. Production of lactic acid from sweet whey permeate concentrates, Process Biochem., 22: 185–188.
Muir, D. D., 1996. Production and use of microbial enzymes for dairy processing J. Soc. Dairy Technol., 49: 24–32
Svendsen A., 2000. Lipase protein engineering, Biochim Biophys Acta 1543 (2): 223-228.
Wilkinson, M. G., 1995. in Cheese – Chemistry, Physics and Microbiology – General Aspects (ed. Fox, P. F.), Chapman and Hall, London, 2nd ed. (1): 523–555.
Ziegler, P., 1999. Determination of the End of Shelf-life for Milk using Weibull Hazard Method, Journal of cereal science 29 (3): 195-204
PCR
Amplify any piece of DNA that we have. Before PCR invented:
- The use of thermostable DNA polymerase
- The development of thermal cyclers or PCR machines
All molecular work since that.
PCR reagent component
- Template (DNA or RNA)
- Two primers
- d-NTPs
- Thermostable DNA polymerase
- PCR buffer
- KCl Tris, MgCl2
Only restriction enzymes
Theoretical: unlimited amounts of DNA
Reasons of result: Competition of complementary daughter strands with primers for re-annealing
Loss of enzyme activity
The number of molecules of polymerase is limiting
Quality of primers.
PRIMERS
Another critical for PCR
Usually 15-30 nucleotides in length
Confirm uniqueness of primers with computer programs
Optimized base pairing at 3’ ends is critical
Random base distribution and average G-C content
Avoid A+T long A+T and G+C rich regions if possible
Minimize internal secondary structure. It is hard to see primer membentuk ikatan dengan dirinya sendiri.
Equal Tm for both primes
Note: look the 3’ ends!!!
Normal primer range: 0.1-1.0 mM
Avoid primer dimer formation (reaksi antar primer sendiri)
Tm (( number of A+T residues)x2oC))+((number of G+C residues)x4oC)
Why 5 degrees lower..because it will be too hot, why? Because for specificity. To force the primer to only bond to the template in the right place (specific product) but to make sure that the hydrogen bond formed.
For long-range PCR, non proofreading + proofreading polymerase are needed to make it possible
PEMILIHAN SEL HELA& MEDIA & STERILISASI AREA KERJA
- Tergantung kebutuhan
- Hela diperoleh dari cell culture collection. Pada online katalog misal di ECACC
- Bisa dipilih media lain, tidak terikat hanya satu macam
- Semua pekerjaan kecuali pengadukan dilakukan dalam safety bench + UV. Setiap alat disemprot dengan alcohol. Tangan disemprot sampai siku. Jangan menyentuhkan ujung pipet pada apapun. Setelah dipakai jangan gunakan pipet yang sama.
- Usahakan membuka tutup T-flask dengan satu tangan. Jangan menggerakkan tangan di atas botol yang terbuka.
- Selalu siapkan vials pembuangan dalam safety bench. Jaga sesedikit mungkin alat-alat di dalamnya
- Tempelkan manual pada dinding safety bench sehingga tidak perlu memegangnya
- Pada saat pemipetan hindari ujung pipet mengenai apapun selain cairan.
PEMBUATAN MEDIUM SEL HELA
- RPIM media + KCl+ pengaduk magnetik dalam 1000 ml beaker
- KCl berfungsi sebagai penjaga pH
- Ambil 225,5 mL media plus buffer + 25 FCS+ 2.5 mL Pen Strep
- Pen/Strep berfungsi sebaga antibiotic agar media steril
- Fungsi FCS untuk menghentikan reaksi-reaksi yang mungkin tejadi agar tidak mengganggu
- Filter larutan, idealnya dengan pompa peristatis tapi bila tidak ada dengan pompa sedot mesin dengan kertas saring umumnya ukuran 0.2 untuk membuang bakteri2 berukuran lebih besar dari 0.2. Lebih kecil ukuran kertas filter lebih baik
PENGHITUNGAN SEL
- Buang medium yang terdapat pada pada T-flask dengan cara ditegakkan, diambil cairannya dengan pipet. Pipet tidak boleh digunakan lagi (posisi sel-sel melekat pada bagian bawah T-flask)
- Sel dicuci dengan PBS. Fungsi PBS:
- Menciptakan keadaan ‘happy’ untuk sel walaupun tanpa ada nutrient
- Mencegah ‘stress’ pada sel saat pencucian
- Tanpa Ca dan Mg karena mereka akan mengaktifkan enzim2 yang mungkin ada dan mengaktivasi sel2 malah mengganggu. Pengadukan dilakukan dengan digoyang kanan-kiri disturb the attachment
- Penambahan trypsin, berguna meng-chelate sel
- Di-tapped agar sel2 terlepas dari dasar T-flask
- Untuk memastikan di-cek di bawah mikroskop
- Biarkan selama 8 menit, tidak terlalu lama karena tripsin dapat membunuh sel
- i. Cheap trypsin conain bad proteins that can kill or
- Penambahan 10% FCS
- Berguna untuk menghentikan reaksi kelatisasi trypsin
- Sentrifuge:
- Bertujuan untuk mengambil hanya sel-sel tanpa bahan-bahan lain termasuk trypsin, FCS
- Sel-sel akan mengendap
- Buang bagian supernatant. Tambahkan lagi 5 ml media komplit
- Perhitungan dengan Hemositometer
- Masukkan 50:50 ke tabung sterile suspense sel + trypan blue solution untuk mewarnai sel.
- Aduk dengan keluar masuk pipet
- Siapkan neubauer chamber dengan menempelkan coverslip sehingga ada gais2 vertikal pelangi untuk memastikan dia menempel. Boleh diberi sedikit uap agar menempel
- Teteskan suspensei sel pada garis coverslip. Lap dengan tissue bila ada cairan di atas tutup untuk menghindari kesalahan perhitungan
- Letakkan chamber di bawah mikroskop, Temukan bentuk seperti di bawah ini, dibatasi 3 garis di luar kotak
| 35 |
|
38 |
| 36 | 35 |
- Maka jumlah sel: (35+36+38+35)/4 x 104 x 2 (factor pengenceran) = 72×104 = 7.2 x105 cell/ ml
- Kita punya 7.2 x 105 cell/ mL, utuk seeding diperlukan 0.5 x 10 5 cell/ ml sebanyak 5 ml sehingga perlu diencerkan
- Dengan rumus V1 M1 = V2 M2 diperoleh:
- (0.5 cell/ml/ 7.2 cell/ ml)x 5 ml = 0.34ml atau 340 µl volume suspend sehingga untuk membuat 5 ml diperlukan penambahan 5000 µl-340 µl = 4660 µl medium.
TRANSFEKSI SEL HELA
- Transfection = Insert naked DNA or plasmid into cells. How to?
- CaPO4 bind to DNA
- Plasmid and molecule attached to CaPO4
- Endositosis
ISOLASI SEL MAKROFAG MENCIT
- Mencit dimatikan dengan mencekik atau dengan kloroform
- Perut dikuliti, disuntikkan media + udara (udara sangat penting dimasukkan untuk membantu pengocokan)
- Perut digoyang-goyang hingga beberapa saat setelah itu cairan media diambil kembali sebanyak kira-kira 5 ml dengan injector yang sama
PASSAGING
Prinsipnya adalah merenew, mengganti medium dengan yang baru
- Cek derajat konfluensinya…(sel2 yang menempel pada dasar flask how much percentage compare to the area size of your microscope lense)
- Detach the cell by trypsin, or by tapping.
- Trypsin hanya bila perlu
- Kita dapatkan loose cells
- Do I have to count? Bila perlu saja
- Rasi Split 1:3 ambil 1/3 bagian dari flask, tambahkan 2/3 medium
According to Barron’s philosophy, English for non-native speakers is a business tool and it has to show dividends immediately. A company that invests in computer technology for its business operations does not wait years before it expects some returns. It is a question of degree. A student needs to listen and understand as much as they need to speak. Inevitably all students want to speak English, but they forget the other half of the formula. Teachers’ aim is to show the students how to communicate their message with the best possible language they can muster.
English for business is a real and living language so teachers should use authentic and current material. Teachers’ text books can be newspapers (British/American) magazines, trade journals, and technical texts, sources from the Internet, leaflets, brochures, company information, radio, and TV and company videos. A text-book is useful for background material whereas authentic material is an ideal source of up to date information. Keep up to date. It’s the professional way.
Teachers should always focus on: the needs of the business people and on specific goals. These are usually identified after a few sessions. Teachers should try to focus their sessions on: things that interest my students, things that the student can relate to, common experiences and things in common to the group. Have a common denominator.
Their sessions are usually based on discussions. They should join corrections, grammar points and new vocabulary in the progress of the discussion. This takes some preparation and business people as students don’t have to fit to their lesson plan. The subjects and topics they choose for each session depend on the interests and needs of the students. This means that they have to be flexible on what I do. There are, however, two topics they make sure they cover, on separate occasions, with each group of new students. These are: problems they have with learning English and mistakes they make when using English.
After all, the Barron’s philosophy wants to make sure teachers make students aware and not self-conscious of their problems and mistakes with English. It also aims to have all my students speak for the majority of the time. Expressing their opinion is important, but asking a why question will help all students express themselves.
Topic-based language learning means that the emphasis of the lessons in language learning is on a subject, a topic or a theme, and the contents of the book are arranged around this topic. It also means that the lesson will include doing work on a topic in class.
Topic based teaching is not the only way to organize teaching, but it is useful, helpful, practical and exciting way to teach either all of the time or some of the time.
There are many benefits of topic-based language learning. First, it is easier to relate the lesson to the experiences and interests of the pupils. Second, the children can associate words, functions, structures and situations with certain topic while we know association can help both understanding and memory. Third, it will bring learners and their needs more into focus. Fourth, it allows teacher to give a personal or local touch to materials depend on the particular class at the particular time. Fifth, the amount of time that the teachers spend can be as long as they like depend on how much time is available and how much material they have.
There are steps to set about topic-based learning. In planning time, teachers should decide at the long term planning stage which topics they are going to work and how long they plan to spend on each topic. In collecting material, once teachers have some idea of possible topics, they should start looking for materials at once. In the functions and situations, once teachers have their topic and collections of connected materials, they should work out which situations and functions of the language they want to concentrate on. When in doubt, it is better to choose the situation before the function. In methods and activities, topic-based work opens up all shorts possibilities. Teachers can choose free activity to lead to much more creative thinking for example. For assessment, the task-based learning gives teachers and the pupils a good opportunity to assess what they have been doing and it is well worth starting in a very casual way.
In summary, I believe learner motivation increases when students are learning about something, rather than just studying language. Topic based learning is particularly appealing in this respect because teachers can use almost any content materials that they feel their students will enjoy.
My Work and Study Highlights 
