Name: preparation of high-quality fermented fish product
Uploaded: 2019-08-23T15:00:00
Description: Watch this Scientific Journal Video about Preparation of High-Quality Fermented Fish Product at JoVE.com

Bu araştırma, Jiangsu Doğal Bilimler Fonu (BK20170185), Jiangsu Balıkçılık Yöneticisi (Y2017-30), Çin Ulusal Doğa Bilimleri Vakfı (NFSC31801575), Çin Tarım Araştırmaları Projesi için ayrılan fon tarafından finansal olarak desteklenmiştir. Sistem (CARS-45-26), Ulusal Birinci Sınıf Gıda Bilimi ve Teknolojisi Disiplin Programı (JUFSTR20180201) ve Yi Tong-Jiangsu Doktora Sonrası Programı.

1. Örnek hazırlama
<ol><li>20 °C'nin altında akan suyun altında dondurulmuş mersin balığı etini eritin. Sonra, temiz, deri kapalı ve küpler halinde balık kesilmiş (2 cm × 2 cm × 1,5 cm). NOT: Hammadde taze veya dondurulmuş mersin balığı eti olabilir.</li>
	<li>10 °C'de 1-3 saat boyunca balık küplerini %6-12 (w/v) tuz çözeltisi ile 1:1 (balık küpü kütlesi çözelti hacmine) oranında karıştırın.</li>
	<li>Marine edilmiş balık küplerini 40-60 °C'de 40-60 °C'de sıcak hava yla yakla...

<ol>
	<li>Riebroy, S., Benjakul, S., Visessanguan, W., Tanaka, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+during+fermentation+and+properties+of+som-fug+produced+from+different+marine+fish.">Changes during fermentation and properties of som-fug produced from different marine fish.</a> Journal of Food Processing and Preservation. 31 (6), 751-770 (2007).</li><li>Xu, Y., Xia, W., Yang, F., Jinmoon, K., Nie, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+fermentation+temperature+on+the+microbial+and+physicochemical+properties+of+silver+carp+sausages+inoculated+with+Pediococcus+pentosaceus.">Effect of fermentation temperature on the microbial and physicochemical properties of silver carp sausages inoculated with Pediococcus pentosaceus.</a> Food Chemistry. 118 (3), 512-518 (2010).</li><li>Cano-Garcia, L., Flores, M., Belloch, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+characterization+and+aromatic+potential+of+Debaryomyces+hansenii+strains+isolated+from+naturally+fermented+sausages.">Molecular characterization and aromatic potential of Debaryomyces hansenii strains isolated from naturally fermented sausages.</a> Food Research International. 52 (1), 42-49 (2013).</li><li>Chen, Q., Liu, Q., Sun, Q., Kong, B., Xiong, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flavour+formation+from+hydrolysis+of+pork+sarcoplasmic+protein+extract+by+a+unique+LAB+culture+isolated+from+Harbin+dry+sausage.">Flavour formation from hydrolysis of pork sarcoplasmic protein extract by a unique LAB culture isolated from Harbin dry sausage.</a> Meat Science. 100, 110-117 (2015).</li><li>Saithong, P., Panthavee, W., Boonyaratanakornkit, M., Sikkhamondhol, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+a+starter+culture+of+lactic+acid+bacteria+in+plaa-som,+a+Thai+fermented+fish.">Use of a starter culture of lactic acid bacteria in plaa-som, a Thai fermented fish.</a> Journal of Bioscience &amp; Bioengineering. 110 (5), 553-557 (2010).</li><li>Zeng, X., Xia, W., Jiang, Q., Guan, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biochemical+and+Sensory+Characteristics+of+Whole+Carp+Inoculated+With+Autochthonous+Starter+Cultures.">Biochemical and Sensory Characteristics of Whole Carp Inoculated With Autochthonous Starter Cultures.</a> Journal of Aquatic Food Product Technology. 24 (1), 52-67 (2015).</li><li>Casaburi, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biochemical+and+sensory+characteristics+of+traditional+fermented+sausages+of+Vallo+di+Diano+(Southern+Italy)+as+affected+by+the+use+of+starter+cultures.">Biochemical and sensory characteristics of traditional fermented sausages of Vallo di Diano (Southern Italy) as affected by the use of starter cultures.</a> Meat Science. 76 (2), 295-307 (2007).</li><li>Gao, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+autochthonous+starter+cultures+on+the+volatile+flavour+compounds+of+Chinese+traditional+fermented+fish+(Suan+yu).">Effect of autochthonous starter cultures on the volatile flavour compounds of Chinese traditional fermented fish (Suan yu).</a> International Journal of Food Science and Technology. 51 (7), 1630-1637 (2016).</li><li>Zeng, X. F., Xia, W. S., Jiang, Q. X., Xu, Y. S., Fan, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+Mixed+Starter+Cultures+to+Flavor+Profile+of+Suanyu+-+A+Traditional+Chinese+Low-Salt+Fermented+Whole+Fish.">Contribution of Mixed Starter Cultures to Flavor Profile of Suanyu - A Traditional Chinese Low-Salt Fermented Whole Fish.</a> Journal of Food Processing and Preservation. 41 (5), 15 (2017).</li><li>Anihouvi, V. B., Sakyi-Dawson, E., Ayernor, G. S., Hounhouigan, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microbiological+changes+in+naturally+fermented+cassava+fish+(Pseudotolithus+sp.)+for+lanhouin+production.">Microbiological changes in naturally fermented cassava fish (Pseudotolithus sp.) for lanhouin production.</a> International Journal of Food Microbiology. 116 (2), 287-291 (2007).</li><li>Corral, S., Salvador, A., Flores, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Salt+reduction+in+slow+fermented+sausages+affects+the+generation+of+aroma+active+compounds.">Salt reduction in slow fermented sausages affects the generation of aroma active compounds.</a> Meat Science. 93 (3), 776-785 (2013).</li><li>Wang, W., Xia, W., Pei, G., Xu, Y., Jiang, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proteolysis+during+fermentation+of+Suanyu+as+a+traditional+fermented+fish+product+of+China.">Proteolysis during fermentation of Suanyu as a traditional fermented fish product of China.</a> International Journal of Food Properties. 20 (90), 29-38 (2017).</li><li>Coventry, M. J., Hickey, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+spices+and+manganese+on+meat+starter+culture+activity.">The effect of spices and manganese on meat starter culture activity.</a> Meat Science. 33 (3), 391-399 (1993).</li><li>Zhu, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+study+on+quality+characteristics+of+pickled+and+fermented+sturgeon+(Acipenser+sinensis)+meat+in+retort+cooking.">Comparative study on quality characteristics of pickled and fermented sturgeon (Acipenser sinensis) meat in retort cooking.</a> International Journal of Food Science &amp; Technology. , (2019).</li><li>Mendonca, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamics+of+the+yeast+flora+in+artisanal+country+style+and+industrial;dry+cured+sausage+(yeast+in+fermented+sausage).">Dynamics of the yeast flora in artisanal country style and industrial;dry cured sausage (yeast in fermented sausage).</a> Food Control. 29 (1), 143-148 (2013).</li><li>Zeng, X. F., Xia, W. S., Jiang, Q. X., Yang, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+autochthonous+starter+cultures+on+microbiological+and+physico-chemical+characteristics+of+Suan+yu,+a+traditional+Chinese+low+salt+fermented+fish.">Effect of autochthonous starter cultures on microbiological and physico-chemical characteristics of Suan yu, a traditional Chinese low salt fermented fish.</a> Food Control. 33 (2), 344-351 (2013).</li><li>Gao, P., Jiang, Q., Xu, Y., Xia, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Esterase+activities+of+autochthonous+starter+cultures+to+increase+volatile+flavor+compounds+in+Chinese+traditional+fermented+fish+(Suan+yu).">Esterase activities of autochthonous starter cultures to increase volatile flavor compounds in Chinese traditional fermented fish (Suan yu).</a> International Journal of Food Properties. 20 (1), 663-672 (2017).</li><li>Wahyuni, S., Holilah, , Asranudin, , Noviyanti, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estimation+of+shelf+life+of+wikau+maombo+brownies+cake+using+Accelerated+Shelf+Life+Testing+(ASLT)+method+with+Arrhenius+model.">Estimation of shelf life of wikau maombo brownies cake using Accelerated Shelf Life Testing (ASLT) method with Arrhenius model.</a> IOP Conference Series Earth and Environmental Science. 122 (1), 12-22 (2018).</li><li>Zeng, X. F., Xia, W. S., Jiang, Q. X., Yang, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+and+microbial+properties+of+Chinese+traditional+low-salt+fermented+whole+fish+product+Suan+yu.">Chemical and microbial properties of Chinese traditional low-salt fermented whole fish product Suan yu.</a> Food Control. 30 (2), 590-595 (2013).</li><li>Yu, D., Jiang, Q., Xu, Y., Xia, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+shelf+life+extension+of+refrigerated+grass+carp+(Ctenopharyngodon+idellus)+fillets+by+chitosan+coating+combined+with+glycerol+monolaurate.">The shelf life extension of refrigerated grass carp (Ctenopharyngodon idellus) fillets by chitosan coating combined with glycerol monolaurate.</a> International Journal of Biological Macromolecules. 101, 448-454 (2017).</li><li>Czerner, M., Tomas, M. C., Yeannes, M. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ripening+of+salted+anchovy+(Engraulis+anchoita):+development+of+lipid+oxidation,+colour+and+other+sensorial+characteristics.">Ripening of salted anchovy (Engraulis anchoita): development of lipid oxidation, colour and other sensorial characteristics.</a> Journal of the Science of Food and Agriculture. 91 (4), 609-615 (2011).</li></ol>

Bu çalışmada, şarap aroması ile yüksek kaliteli fermente balık ürününün üretimi için yeni bir teknik ve duyusal özellikleri için testler sağlanmıştır. Bu tekniğin temel süreçleri marine, kurutma, fermantasyon ve sterilizasyon vardır. Marine işleminde tuz konsantrasyonu, sıcaklık ve zaman balığın dokusal özelliklerini etkiler. Tuz konsantrasyonunun artmasıyla (0-%12, w/v) ve marine süresinin uzaması (0-2,5 saat) ile ürünün sertliği ve çiğnenme oranı kademeli olarak artar; ancak 2 sa...

Çin'de mevcut ticari marine balık ürünü ağır tuzlu tat, yetersiz şarap aroması, kötü elastikiyet ve soluk renk sorunu vardır, hangi tüketicilere kabul edilebilirlik azaltır. Bu nedenle, şarap aroması ile yüksek kaliteli balık eti ürün için yeni bir teknik oluşturulmalı ve optimize edilmelidir.
Son yıllarda, et ve balık modern fermantasyon teknikleri uygulama dahafazla araştırmacı 1,2,3,4dikkat çekti . Başlangıç kültürlerinin et ve balık lara aşılanmasıyla gıda güvenliği artırıldı, işleme süresi kısaldı; ve ürün duyusal özellikleri değiştirilmiştir. Saithong ve ark. doğal plaa-som gelen5 izole lactobacillus bakteri (LAB) ve bir başlangıç kültürü olarak bu LAB kullanılan, hangi yüksek asitlik indüklenen ve patojenik bakteriler bastırılmış. Zeng ve ark.6 otokton başlangıç kültürleri ile aşılama fermantasyon süresini azalttığını ve örneklerin duyusal özelliklerini iyileştirdiğini bildirdi. Casaburi ve ark. 7, mikrobiyal başlangıç kültürlerinin kullanımının fermente etlerde aroma gelişimini etkilediğini iddia ettiler. Bu başlangıç kültürlerde, S. cerevisiae alkollü fermantasyon ile şarap aroması üretebilir ve aynı zamanda ürün diğer geliştirilmiş organoleptik nitelikleri verebilir. Bu nedenle, S. cerevisiae şarap-aroma ürünleri8,9,10 ve şarap-aroma balık ürün S. cerevisiaetarafından yapılabilir için uygun bir başlangıç kültürüdür.
Şarap-aroma balık ürün yapma sürecinde, et ve balık dokusu tuz içeriği, su içeriği, pH, protein denatürasyonu, vb etkilenebilir Bu nedenle, marine, kurutma, fermantasyon ve sterilizasyon tüm doku özelliklerini etkileyebilir. Lezzet ve tat oluşumu karmaşıktır ve yüksek karbonhidrat, protein ve lipidlerin hidrolizi ile ilgili olduğu için, özellikle marine ve fermantasyon etkilenir, ve hafif lipid oksidasyonu11,12. Ayrıca baharat 13 eklenmesindende etkilenebilir. Renk gelişimi için, Fermantasyon ve sterilizasyon sürecinde yer alan Maillard reaksiyonu oluşur10.
Bu makale, balık işleme endüstrisinin gelişimi için büyük önem taşıyan şarap aroması ile fermente balık ürününün sanayileşmesi için teknik destek sağlayabilir. Bu teknik artan proteoliz (daha serbest amino asitler ve TCA çözünür peptidler) ile ürünün tadını artırabilir, alkoller (etanol, 1-oksen-3-ol, 2-metil-1-propanol ve 3-metil-1-butanol), esterler (etil asetat) ve özellikle lezzet değiştirmek aldehitler (nonanal, 3-metilbutanal ve benzaldehit), yüksek sertlik, yaylılık, gumminess ve chewiness tarafından mouthfeel artırmak ve daha çekici russet renk vermek ve parlak bir yüzey14. Ayrıca tüketicilere kolaylık sağlar, çünkü ürün oda sıcaklığında saklanabilir. Diğer önceki çalışmalarda açıklandığı gibi15,16,17, S. cerevisiae ile fermantasyon da önemli ölçüde diğer et veya balık ürünlerinde organoleptik nitelikleri geliştirmek için kanıtlanmıştır.
Bu tanıtılan protokol de ot sazan, gümüş sazan, siyah sazan, bighead sazan, morina, somon, vb gibi balık diğer türler, uygulanabilir dikkati çekiyor. Balık ürünlerinin yüksek kalitesi için, işleme olmadan balık, taze balık, buz balık veya dondurulmuş balık az 1 yıl için saklanan gibi kullanılmalıdır. Ayrıca, geniş lipid oksidasyonu tatsız bir lezzet getirirken hafif lipid oksidasyonu lezzeti artırabileceğinden, daha az yağ içeren balıklar tercih edilir veya yağsız balık önerilir.

<table>
	<tbody>
		<tr>
			<td>2,4,6-trimethylpyridine</td>
			<td>Tokyo Chemical Industry Co., Ltd.</td>
			<td>Purity 98%</td>
			<td></td>
		</tr>
		<tr>
			<td>Colorimeter</td>
			<td>Hunterlab</td>
			<td>UltraScan Pro1166</td>
			<td></td>
		</tr>
		<tr>
			<td>DB-WAX column</td>
			<td>Agilent</td>
			<td>30 m &#215; 0.25 mm &#215; 0.25&#956;m</td>
			<td></td>
		</tr>
		<tr>
			<td>Digital pH meter</td>
			<td>Mettler toledo Instrument (Shanghai) Co., Ltd.</td>
			<td>DELTA-320</td>
			<td></td>
		</tr>
		<tr>
			<td>Drying oven</td>
			<td>Shanghai Yiheng Scientific Instrument Co., Ltd.</td>
			<td>DHG-9070A</td>
			<td></td>
		</tr>
		<tr>
			<td>Frozen sturgeon</td>
			<td>Huada Marine Industry Group Co., Ltd</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>Gas chromatograph-mass spectrometer</td>
			<td>Thermo Fisher Scientific</td>
			<td>TSQ Quantum XLS</td>
			<td></td>
		</tr>
		<tr>
			<td>Humidities incubator</td>
			<td>Shanghai Yiheng Scientific Instrument Co., Ltd.</td>
			<td>LHS-250HC-II</td>
			<td></td>
		</tr>
		<tr>
			<td>Saccharomyces cerevisiae</td>
			<td>Angel Yeast Co., Ltd</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>Spices</td>
			<td>Auchan Supermarket</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterilization pot</td>
			<td>Longqiang Machinery Technology Co., Ltd.</td>
			<td>RHS-03-700</td>
			<td></td>
		</tr>
		<tr>
			<td>Supelco</td>
			<td>Sigma</td>
			<td>65&#956;m, PDMS/DVB</td>
			<td></td>
		</tr>
		<tr>
			<td>Texture analyzer</td>
			<td>Stable Micro Systems, Ltd.</td>
			<td>TA-XT2i</td>
			<td></td>
		</tr>
		<tr>
			<td>Vacuum package machine</td>
			<td>Quanzhou Yiminxin Electromechanical Co., Ltd.</td>
			<td>YMX-958-10L</td>
			<td></td>
		</tr>
	</tbody>
</table>

preparation of high-quality fermented fish product

Bu protokol, mersin balığı(Aquilaria sinensis)et ürünü ile sanayileşmiş fermente balık ürününün hazırlanması için bir yöntem sağlamaktadır. Prosedürler şunlardır: (1) decapitation, evisceration, skinning-off, temizlik ve kesme dahil olmak üzere çiftlik mersin balığı ön tedavi; (2) 6-12% (w / v) tuz çözeltisi (1:1, çözelti hacmine balık küpü kütlesi) balık küpleri marine; (3) sıcak hava (40-60 °C) veya vakum ile% 50-60 su içeriğine balık küpleri kurutma; (4) balık küpleri ile aşılanan fermantasyon 0.4-1.6% (w / w) S. cerevisiae lezzet çözeltisi balık küpleri ve 6-10 saat için 25-35 °C'de fermantasyon; (5) marine ve fermantasyon çözümleri ile vakum paketleri balık küpleri sızdırmazlık; (6) 10-20 dakika boyunca 115-121 °C'de sterilize edilir. Bu yöntemle hazırlanan mersin balığı eti ürünü yumuşak ve kalın lezzetli bir tadı vardır, çeşitli türleri ve alkoller ve balıklardan gelen tuzlu ve tatsız kokuyu maskeleyebilecek esterler gibi uçucu lezzet bileşikleri büyük miktarlarda vardır, orta tuz vardır içerik ama yüksek yaylılık, gumminess ve chewiness gibi iyi doku özellikleri ve parlak russet renk ve çekici bir görünüme sahiptir. Bu yeni teknik aynı zamanda oda sıcaklığında saklanabilir uygun balık çerez gıdalar sağlamak için diğer balık işleme uygulanabilir. Hem deniz hem de tatlı su balıkları için uygundur.

Uygun tuz konsantrasyonu, marine süresi ve sıcaklığı ürünün dokusal kalitesini daha iyi hale getirir. En iyi marine koşulları aşağıdaki gibidir: marine çözeltisindeki tuz konsantrasyonu %8;; 2 saat marine süresi; ve 10 °C'lik marine sıcaklığı. Bkz. Şekil 1.
Kurutma işleminde, son nem içeriği ve kurutma sıcaklığı doku ve duyusal kaliteyi etkileyebilir. En iyi kurutma koşulları şunlardır: kurutulmuş balık küplerinin %55 nem içeri...

Watch this Scientific Journal Video about Preparation of High-Quality Fermented Fish Product at JoVE.com

Preparation of High-Quality Fermented Fish Product

Protokolün amacı Saccharomyces cerevisiaeaşılama dayalı bir sanayileşmiş balık fermantasyon tekniği sağlamaktır.

Yüksek Kaliteli Fermente Balık Ürününün Hazırlanması

This protocol provides a method for preparation of industrialized fermented fish product with sturgeon (Aquilaria sinensis) meat product. The procedures ...

Bu prosedür, şarap aroması ile fermente balık ürün yüksek kaliteli üretir ve sanayi üretiminde uygulanabilir. Bu teknik balık ürün çekici bir görünüm, benzersiz lezzet, lezzetli tat ve iyi doku özellikleri verir. Ve bu ürün en az bir yıl oda sıcaklığında saklanabilir.Bu yordamı başlatmak için, çözülme, temiz ve cilt dondurulmuş mersin balığı eti metin protokolünde belirtildiği gibi. Küpler halinde balık kesin. Daha sonra, bir ila üç saat boyunca 10 santigrat derece bire bir oranında% 6-12 tuz çözeltisi ile balık küpleri karıştırın.Daha sonra, paslanmaz çelik ızgara plaka üzerine marine balık küpleri koymak ve 6 ila 10 saat boyunca 40 ve 60 santigrat derece arasında bir sıcaklıkta sıcak hava ile kurulayın. Baharat sıvısını hazırlamak için, 1,2 birim taze soğan, bir birim zencefil, 0,6 ünite yıldız anason, 0,6 adet rezene, 0,3 adet yeşil çay ve 40 ünite su ile 0,3 birim biber içeren baharatları karıştırın. Yarım saat kaynatın ve filtrat toplamak.Lezzet çözeltisi hazırlamak için, burada gösterilen kitle fraksiyonları baharat sıvı, sarı şarap, beyaz şarap, şeker kamışı, monosodyum glutamat ve sirke karıştırın. Bundan sonra, bire bir oranında 0,4 ila% 1,6 saccharomyces cerevisiae aşılama, lezzet çözeltisi ile kurutulmuş balık küpleri karıştırın. Karışımı bir kapta kapatın ve 25 ila 35 santigrat derece arasında bir sıcaklıkta altı ila 10 saat boyunca fermente edin.Fermente balık küplerini marine ve fermantasyon çözümleriyle bir vakum paketine yerleştirin ve vakum paketini 0,085 megapascal, 3,6 saniyelik bir sızdırmazlık süresi ve 5,5 saniyelik bir soğutma süresiyle kapatın. Daha sonra 10 ila 20 dakika boyunca 115 ve 121 santigrat derece arasında bir sterilizasyon sıcaklığı kullanarak paketlenmiş balık küpleri sterilize. Bu çalışmada yüksek kaliteli fermente balık ürünü hazırlanmıştır.Uygun bir tuz konsantrasyonu kullanarak, marine süresi ve sıcaklık ürünleri dokusal kalite daha iyi hale getirir. En iyi marine koşulları% 8 tuz konsantrasyonu ve marine çözeltisi, iki saatlik bir marine süresi gibi görünüyordu. Ve 10 santigrat derecelik bir marine sıcaklığı.Kurutma işleminde, son nem içeriği ve kurutma sıcaklığı doku ve sensör kalitesini etkileyebilir. En iyi kurutma koşulları% 55 kurutulmuş balık küpleri ve 50 santigrat derece kurutma sıcaklığı bir nem içeriği gibi görünüyordu. Fermantasyon sürecinde, sakaromyces cerevisiae eklenmesi, sıcaklık, ve zaman tüm etkisi tat, lezzet, renk, ve balık doku özellikleri.Başarılı fermente balık tat, lezzet, renk ve doku, uyumlu tat, fermantif koku, şarap, russet renk, parlak yüzey, karamel ve al dente doku sonuçlanan yüksek puanları vardır. Başarısız balık ürün tat, lezzet, renk ve doku kötü tat, acı, ekşilik, balıklık, soluk renk, pürüzlü yüzey, kaba mouthfeel ve bozulmuş doku sonuçlanan düşük puanları vardır. Uygun sterilizasyon ve yoğunluk 4.5 dakika gibi görünüyor.Ve uygun bir sterilizasyon sıcaklığı 121 santigrat derece gibi görünüyor. Depolama hususları daha sonra metin protokolünde belirtildiği şekilde hesaplanır ve belirlenir. Ve raf ömrü için tahmin 264 gün 25 santigrat derece veya 205 gün 35 santigrat derece.Bu işlemi gerçekleştirirken hem fermantasyonun hem de sterilizasyon adımlarının bu tekniğin başarısı için hayati önem taşıdığını belirtmek önemlidir. Bu prosedürü öğrendikten sonra, diğer balık türlerinin fermantasyon kullanılabilir. Gelecekte, diğer suşları ile fermantasyon araştırılabilir.Lactobacillus, staphylococcus, vesaire gibi. bölgelerde, balık ürünlerini sterilize ederken önlem almayı unutmayın, çünkü son derece yüksek sıcaklık zarar verebilir.

preparation-of-high-quality-fermented-fish-product

Research

JoVE Journal

Biochemistry

Preparation of Homogeneous MALDI Samples for Quantitative Applications

This protocol demonstrates a simple sample preparation to reduce spatial heterogeneity in ion signals during matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The heterogeneity of ion signals is a severe problem in MALDI, which results in poor data reproducibility and makes MALDI unsuitable for quantitative analysis. By regulating sample plate temperature during sample preparation, thermal-induced hydrodynamic flows inside droplets of sample solution are able to reduce the heterogeneity problem. A room-temperature sample preparation chamber equipped with a temperature-regulated copper base block that holds MALDI sample plates facilitates precise control of the sample drying condition. After drying of sample droplets, the temperature of sample plates is returned to room temperature and removed from the chamber for subsequent mass spectrometric analysis. The areas of samples are examined with MALDI-imaging mass spectrometry to obtain the spatial distribution of all components in the sample. In comparison with the conventional dried-droplet method that prepares samples under ambient conditions without temperature control, the samples prepared with the method demonstrated herein show significantly better spatial distribution and signal intensity. According to observations using carbohydrate and peptide samples, decreasing substrate temperature while maintaining the surroundings at ambient temperature during the drying process can effectively reduce the heterogeneity of ion signals. This method is generally applicable to various combinations of samples and matrices.

A protocol for reducing spatial heterogeneities of ion signals in MALDI mass spectrometry by regulating substrate temperature during sample drying processes is demonstrated.

Kantitatif Uygulamalar için Homojen MALDI Örneklerin Hazırlanması

This protocol demonstrates a simple sample preparation to reduce spatial heterogeneity in ion signals during matrix-assisted laser desorption/ionization ...

Biyokimya

A Filtration-based Method of Preparing High-quality Nuclei from Cross-linked Skeletal Muscle for Chromatin Immunoprecipitation

Chromatin immunoprecipitation (ChIP) is a powerful method to determine protein binding to chromatin DNA. Fiber-rich skeletal muscle, however, has been a challenge for ChIP due to technical difficulty in isolation of high-quality nuclei with minimal contamination of myofibrils. Previous protocols have attempted to purify nuclei before cross-linking, which incurs the risk of altered DNA-protein interaction during the prolonged nuclei preparation process. In the current protocol, we first cross-linked the skeletal muscle tissue collected from mice, and the tissues were minced and sonicated. Since we found that ultracentrifugation was not able to separate nuclei from myofibrils using cross-linked muscle tissue, we devised a sequential filtration procedure to obtain high-quality nuclei devoid of significant myofibril contamination. We subsequently prepared chromatin by using an ultrasonicator, and ChIP assays with anti-BMAL1 antibody revealed robust circadian binding pattern of BMAL1 to target gene promoters. This filtration protocol constitutes an easily applicable method to isolate high-quality nuclei from cross-linked skeletal muscle tissue, allowing consistent sample processing for circadian and other time-sensitive studies. In combination with next-generation sequencing (NGS), our method can be deployed for various mechanistic and genomic studies focusing on skeletal muscle function.

We present a filtration-based protocol to isolate high-quality nuclei from cross-linked mouse skeletal muscle wherein we removed the need for ultracentrifugation, making it easily applicable. We show that chromatin prepared from the nuclei is suitable for chromatin immunoprecipitation and likely chromatin immunoprecipitation sequencing studies.

Kromatin İmmunopresipitasyon için Çapraz Bağlı İskelet Kasında Yüksek Kaliteli Nükleerlerin Hazırlanması İçin Filtrasyona Dayalı Bir Yöntem

Chromatin immunoprecipitation (ChIP) is a powerful method to determine protein binding to chromatin DNA. Fiber-rich skeletal muscle, however, has been a ...

Biosynthesis of a Flavonol from a Flavanone by Establishing a One-pot Bienzymatic Cascade

Flavonols are a major subclass of flavonoids with a variety of biological and pharmacological activities. Here, we provide a method for the in vitro enzymatic synthesis of a flavonol. In this method, Atf3h and Atfls1, two key genes in the biosynthetic pathway of the flavonols, are cloned and overexpressed in Escherichia coli. The recombinant enzymes are purified via an affinity column and then a bienzymatic cascade is established in a specific synthetic buffer. Two flavonols are synthesized in this system as examples and determined by TLC and HPLC/LC/MS analyses. The method displays obvious advantages in the derivation of flavonols over other approaches. It is time- and labor-saving and highly cost-effective. The reaction is easy to be accurately controlled and thus scaled up for mass production. The target product can be purified easily due to the simple components in the system. However, this system is usually restricted to the production of a flavonol from a flavanone.

The derivation of a flavonol is crucial for its application in healthcare and the food industry. Here, we provide a detailed protocol for the biosynthesis of a flavonol from a flavanone and discuss the crucial steps and its advantages over other approaches.

Bir Flavanone bir Flavonol Biyosentezi Bir-pot Bienzymatik Cascade Kurarak

Flavonols are a major subclass of flavonoids with a variety of biological and pharmacological activities. Here, we provide a method for the in vitro ...

Obtaining High-Quality Transcriptome Data from Cereal Seeds by a Modified Method for Gene Expression Profiling

The characterization of gene expression is dependent on RNA quality. In germinating, developing and mature cereal seeds, the extraction of high-quality RNA is often hindered by high starch and sugar content. These compounds can reduce both the yield and the quality of the extracted total RNA. The deterioration in quantity and quality of total RNA can subsequently have a significant impact on the downstream transcriptomic analyses, which may not accurately reflect the spatial and/or temporal variation in the gene expression profile of the samples being tested. In this protocol, we describe an optimized method for extraction of total RNA with sufficient quantity and quality to be used for whole transcriptome analysis of cereal grains. The described method is suitable for several downstream applications used for transcriptomic profiling of developing, germinating, and mature cereal seeds. The method of transcriptome profiling using a microarray platform is shown. This method is specifically designed for gene expression profiling of cereals with described genome sequences. The detailed procedure from microarray handling to final quality control is described. This includes cDNA synthesis, cRNA labelling, microarray hybridization, slide scanning, feature extraction, and data quality validation. The data generated by this method can be used to characterize the transcriptome of cereals during germination, in various stages of grain development, or at different biotic or abiotic stress conditions. The results presented here exemplify high-quality transcriptome data amenable for downstream bioinformatics analyses, such as the determination of differentially expressed genes (DEGs), characterisation of gene regulatory networks, and conducting transcriptome-wide association study (TWAS).

A method for transcriptome profiling of cereals is presented. The microarray-based gene expression profiling starts with the isolation of high-quality total RNA from cereal grains and continues with the generation of cDNA. After cRNA labelling and microarray hybridization, recommendations are given for signal detection and quality control.

Gen Ekspresyonu Profilleme için Modifiye Yöntemi ile Tahıl Tohumlarından Yüksek Kaliteli Transkripsiyon Verilerinin Elde Edilmesi

The characterization of gene expression is dependent on RNA quality. In germinating, developing and mature cereal seeds, the extraction of high-quality ...

Single Cell Analysis Of Transcriptionally Active Alleles By Single Molecule FISH

Gene transcription is an essential process in cell biology, and allows cells to interpret and respond to internal and external cues. Traditional bulk population methods (Northern blot, PCR, and RNAseq) that measure mRNA levels lack the ability to provide information on cell-to-cell variation in responses. Precise single cell and allelic visualization and quantification is possible via single molecule RNA fluorescence in situ hybridization (smFISH). RNA-FISH is performed by hybridizing target RNAs with labeled oligonucleotide probes. These can be imaged in medium/high throughput modalities, and, through image analysis pipelines, provide quantitative data on both mature and nascent RNAs, all at the single cell level. The fixation, permeabilization, hybridization and imaging steps have been optimized in the lab over many years using the model system described herein, which results in successful and robust single cell analysis of smFISH labeling. The main goal with sample preparation and processing is to produce high quality images characterized by a high signal-to-noise ratio to reduce false positives and provide data that are more accurate. Here, we present a protocol describing the pipeline from sample preparation to data analysis in conjunction with suggestions and optimization steps to tailor to specific samples.&#160;

Single molecule RNA fluorescence in situ hybridization (smFISH) is a method to accurately quantify levels and localization of specific RNAs at the single cell level. Here, we report our validated lab protocols for wet-bench processing, imaging and image analysis for single cell quantification of specific RNAs.

Transkripsiyonel Aktif Alellerin Tek Molekül FISH ile Tek Hücre Analizi

Gene transcription is an essential process in cell biology, and allows cells to interpret and respond to internal and external cues. Traditional bulk ...

Nano-Differential Scanning Fluorimetry for Screening in Fragment-based Lead Discovery

Thermal shift assays (TSAs) examine how the melting temperature (Tm) of a target protein changes in response to changes in its environment (e.g., buffer composition). The utility of TSA, and specifically of nano-Differential Scanning Fluorimetry (nano-DSF), has been established over the years, both for finding conditions that help stabilize a specific protein and for looking at ligand binding by monitoring changes in the apparent Tm. This paper presents an efficient screening of the Diamond-SGC-iNEXT Poised (DSi-Poised) fragment library (768 compounds) by the use of nano-DSF, monitoring Tm to identify potential fragment binding. The prerequisites regarding protein quality and concentration for performing nano-DSF experiments are briefly outlined followed by a step-by-step protocol that uses a nano-liter robotic dispenser commonly used in structural biology laboratories for preparing the required samples in 96-well plates. The protocol describes how the reagent mixtures are transferred to the capillaries needed for nano-DSF measurements. In addition, this paper provides protocols to measure thermal denaturation (monitoring intrinsic tryptophan fluorescence) and aggregation (monitoring light back-scattering) and the subsequent steps for data transfer and analysis. Finally, screening experiments with three different protein targets are discussed to illustrate the use of this procedure in the context of lead discovery campaigns. The overall principle of the method described can be easily transferred to other fragment libraries or adapted to other instruments.

Monitoring changes in the melting temperature of a target protein (i.e., thermal shift assay, TSA) is an efficient method for screening fragment libraries of a few hundred compounds. We present a TSA protocol implementing robotics-assisted nano-Differential Scanning Fluorimetry (nano-DSF) for monitoring intrinsic tryptophan fluorescence and light back-scattering for fragment screening.

Parça Bazlı Kurşun Keşfinde Tarama için Nano-Diferansiyel Tarama Florüretrisi

Thermal shift assays (TSAs) examine how the melting temperature (Tm) of a target protein changes in response to changes in its environment (e.g., buffer ...

Preparation of High-Temperature Sample Grids for Cryo-EM

The sample grids for cryo-electron microscopy (cryo-EM) experiments are usually prepared at a temperature optimal for the storage of biological samples, mostly at 4 &#176;C and occasionally at room temperature. Recently, we discovered that the protein structure solved at low temperature may not be functionally relevant, particularly for proteins from thermophilic archaea. A procedure was developed to prepare protein samples at higher temperatures (up to 70 &#176;C) for cryo-EM analysis. We showed that the structures from samples prepared at higher temperatures are functionally relevant and temperature dependent. Here we describe a detailed protocol for preparing sample grids at high temperature, using 55 &#176;C as an example. The experiment made use of a vitrification apparatus modified using an additional centrifuge tube, and samples were incubated at 55 &#176;C. The detailed procedures were fine-tuned to minimize vapor condensation and obtain a thin layer of ice on the grid. Examples of successful and unsuccessful experiments are provided.

This paper provides a detailed protocol for preparing sample grids at temperatures as high as 70 &#176;C, prior to plunge freezing for cryo-EM experiments.

Cryo-EM için Yüksek Sıcaklık Numune Izgaralarının Hazırlanması

The sample grids for cryo-electron microscopy (cryo-EM) experiments are usually prepared at a temperature optimal for the storage of biological samples, ...

The Extraction of Liver Glycogen Molecules for Glycogen Structure Determination

Liver glycogen is a hyperbranched glucose polymer that is involved in the maintenance of blood sugar levels in animals. The properties of glycogen are influenced by its structure. Hence, a suitable extraction method that isolates representative samples of glycogen is crucial to the study of this macromolecule. Compared to other extraction methods, a method that employs a sucrose density gradient centrifugation step can minimize molecular damage. Based on this method, a recent publication describes how the density of the sucrose solution used during centrifugation was varied (30%, 50%, 72.5%) to find the most suitable concentration to extract glycogen particles of a wide variety of sizes, limiting the loss of smaller particles. A 10 min boiling step was introduced to test its ability to denature glycogen degrading enzymes, thus preserving glycogen. The lowest sucrose concentration (30%) and the addition of the boiling step were shown to extract the most representative samples of glycogen.

An optimal sucrose concentration was determined for the extraction of liver glycogen using sucrose density gradient centrifugation. The addition of a 10 min boiling step to inhibit glycogen-degrading enzymes proved beneficial.

Glikojen Yapısının Belirlenmesi için Karaciğer Glikojen Moleküllerinin Ekstraksiyonu

Liver glycogen is a hyperbranched glucose polymer that is involved in the maintenance of blood sugar levels in animals. The properties of glycogen are ...

Preparation of siRNA Mixture for Mitochondrial DNA Synthesis

To begin, prepare a 140 nano molar mix of small interfering RNA or siRNA by diluting the siRNA in optimum medium. Add the five microliters of the diluted mix to the wells of a black 384 well cell culture microplate. Prepare the appropriate amount of RNAiMAX transfection reagent solution in optimum medium and add 10 microliters to each well of the 384 well plate with a reagent dispenser.

Mitokondriyal DNA Sentezi için SiRNA Karışımının Hazırlanması

In Situ Hücresel Kriyotomografi için Elektron Mikroskobu Izgaralarının Hazırlanması

Sample Preparation for In Situ Cryotomography of Mammalian Cells

In situ cellular cryotomography is a powerful technique for studying complex objects in their native frozen-hydrated cellular context, making it highly relevant to cellular biology and virology. The potential of combining cryotomography with other microscopy modalities makes it a perfect technique for integrative and correlative imaging. However, sample preparation for in situ cellular tomography is not straightforward, as cells do not readily attach and stretch over the electron microscopy grid. Additionally, the grids themselves are fragile and can break if handled too forcefully, resulting in the loss of imageable areas. The geometry of tissue culture dishes can also pose a challenge when manipulating the grids with tweezers. Here, we describe the tips and tricks to overcome these (and other) challenges and prepare good-quality samples for in situ cellular cryotomography and correlative imaging of adherent mammalian cells. With continued advances in cryomicroscopy technology, this technique holds enormous promise for advancing our understanding of complex biological systems.

Yazar Spotlight: Gelişmiş Kriyoelektron Tomografisi için Izgara Hazırlığını Optimize Etme 

This method provides an accessible and flexible protocol for the preparation of electron microscopy (EM) grids for in situ cellular cryotomography and correlative light and electron microscopy (CLEM).

Memeli Hücrelerinin İn Situ Kriyotomografisi için Numune Hazırlama

In situ cellular cryotomography is a powerful technique for studying complex objects in their native frozen-hydrated cellular context, making it highly ...