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

이 연구는 장쑤 자연 과학 기금 (BK20170185), 장쑤 수산 관리자 (Y2017-30), 중국 국립 자연 과학 재단 (NFSC31801575), 중국 농업 연구에 대한 귀한 기금에 의해 재정적으로 지원되었습니다. 시스템 (CARS-45-26), 식품 과학 기술의 국가 일류 훈련 프로그램 (JUFSTR20180201), 그리고 이동 장수 박사 과정.

1. 견본 준비
<ol><li>20 °C 이하의 흐르는 물 에서 냉동 철갑상어 고기를 해동하십시오. 그런 다음, 깨끗한 피부떨어져 큐브 (2cm × 2cm × 1.5 cm)로 물고기를 잘라. 참고 : 원료는 신선하거나 냉동 된 철갑상어 고기 일 수 있습니다.</li>
	<li>생선 큐브를 1-1%(생선 큐브 질량에서 용액 부피)의 속도로 6-12%(v) 소금 용액과 1-3시간 동안 10°C에서 섞습니다.</li>
	<li>양념된 생선 큐브를 스테인리스 스틸 그리딩...

<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>

이 연구에서는 와인 향이 나는 고품질 발효 생선 제품을 생산하기 위한 새로운 기술과 감각 적 특성에 대한 테스트가 제공됩니다. 이 기술의 주요 공정은 양념, 건조, 발효 및 살균입니다. 양념 과정에서 소금 농도, 온도 및 시간은 모두 물고기의 텍스처 특성에 영향을 미칩니다. 소금 농도 (0-12 %, w / v)의 증가와 양념 시간 (0-2.5 h)의 연장으로 제품의 경도와 쫄깃함이 점차 증가합니다. 그러나 2 시간...

중국의 현재 상업 양념 생선 제품은 무거운 짠 맛, 부족한 와인 향기, 불쌍한 탄성 및 옅은 색의 문제가있어 소비자의 수용성을 감소시입니다. 따라서 와인 향이 나는 고품질 생선 고기 제품에 대한 새로운 기술을 만들고 최적화해야 합니다.
최근에는 육류와 생선에 현대 발효 기술의 적용이 점점 더 많은연구자 1, 2,3,4에서주목받고 있다. 육류와 생선에 스타터 배양을 접종함으로써 식품 안전이 향상되었으며 가공 시간이 단축되었습니다. 및 제품 감각 특성이 수정되었습니다. Saithong 외.5 천연 플라솜에서 분리 된 유산 균 박테리아 (LAB) 스타터 배양으로이 LAB을 사용, 이는 높은 산도를 유도하고 병원성 박테리아를 억제. Zeng et al. 6은 자가 착동 배양물로 접종하여 발효 시간을 단축하고 시료의 감각 적 특성을 개선했다고 보고했다. Casaburi 외7 미생물 스타터 배양물의 사용은 발효 육류의 아로마 의 발달에 영향을 미친다고 주장했다. 이러한 스타터 문화에서, S. cerevisiae 알코올 발효에 의해 와인 향기를 생산할 수 있으며 또한 제품 다른 향상 된 관능 적 자질을 줄 수 있습니다. 따라서, S. 세레비시아는 와인-아로마 제품 8,9,10 및 와인-아로마 어패류 제품에 적합한 스타터 문화로 S. cerevisiae에의해 만들어질 수 있다.
와인 아로마 생선 제품을 만드는 과정에서 고기와 생선의 질감은 소금 함량, 수분 함량, pH, 단백질 변성 등에 의해 영향을받을 수 있습니다. 따라서 양념, 건조, 발효 및 살균은 모두 질감 특성에 영향을 줄 수 있습니다. 맛과 맛의 형성은 복잡하고 주로 양념 및 발효에 의해 영향을 받는데, 이는 탄수화물, 단백질 및 지질의 가수분해, 및 경질 지질 산화와 밀접한 관련이 있기 때문이다 11,12. 그것은 또한 향신료13의추가에 의해 영향을 받을 수 있습니다. 색상의 개발을 위해, 발효 및 살균 (10)의 과정에 관여하는Maillard 반응이 발생한다.
이 기사는 와인 향이 나는 발효 어류 제품의 산업화를 위한 기술 지원을 제공할 수 있으며, 이는 어류 가공 산업의 발전에 큰 의미가 있습니다. 이 기술은 증가 된 프로테오 용해 (더 많은 자유 아미노산 및 TCA 수용성 펩티드)에 의해 제품의 맛을 향상시킬 수, 알코올 (에탄올, 1-옥텐-3-올, 2 메틸-1-프로판및 3 메틸-1-부탄올), 에스테르 (에틸 아세테이트)에 의해 주로 맛을 수정 알데히드(nonanal, 3-메틸부탄 및 벤잘데히드)는 더 높은 경도, 탄력, 구민 및 쫄깃함으로 입맛을 증가시키고, 더 매력적인 러셋 색상을 더 하여 밝은 표면14를준다. 또한 제품이 실온에서 보관할 수 있기 때문에 소비자의 편의를 제공합니다. 다른 이전 연구에서 설명한 바와 같이15,16,17, S. 세레비시아와 함께 발효하는 것은 또한 다른 육류 또는 생선 제품의 관능적 특성을 현저히 향상시키는 것으로 입증되었다.
도입 된 프로토콜은 잔디 잉어, 은 잉어, 검은 잉어, 큰 머리 잉어, 대구, 연어 등과 같은 다른 종류의 물고기에도 적용 될 수 있다는 점에 주목할 필요가 있습니다. 생선 제품의 고품질의 경우, 가공하지 않은 생선, 신선한 생선, 얼음에 있는 생선 또는 냉동 생선과 같이 1년 미만 보관된 생선을 사용해야 합니다. 게다가, 온화한 지질 산화는 풍미를 강화할 수 있기 때문에 광대한 지질 산화는 불쾌한 맛을 가져오고, 더 적은 지방을 가진 물고기는 바람직하거나 skimmed 물고기가 좋습니다.

<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

이 프로토콜은철갑상어(Aquilaria sinensis)육류 제품으로 공업화된 발효 생선 제품을 제조하는 방법을 제공한다. 절차는 다음과 이었다: (1) 참수 포함 하 여 농장 된 외과 의 전처리, evisceration, 스키닝 오프, 청소 및 절단; (2) 6-12 % (v) 소금 용액 (1 : 1, 용액 부피에 물고기 큐브 질량)에 생선 큐브를 양념; (3) 뜨거운 공기 (40-60 °C) 또는 진공에 의해 50-60 %의 수분 함량으로 생선 큐브를 건조; (4) 0.4-1.6%(w/w)의 생선 큐브를 접종하는 발효는 6-10시간 동안 25-35°C에서 발효하는 풍미 용액에 0.4-1.6%(w/w)의 세레비시아; (5) 양념 및 발효 용액으로 진공 패키지에 생선 큐브를 밀봉; (6) 115-121 °C에서 10-20 분 동안 살균. 이 방법으로 제조 된 철갑상어 고기 제품은 부드럽고 두껍고, 다양한 유형과 생선의 곰팡이와 불쾌한 냄새를 가릴 수있는 알코올 및 에스테르와 같은 휘발성 맛 화합물이 다량 있으며 적당한 소금을 가지고 있습니다. 내용이 많지만 탄력이 높고, 구미와 쫄깃함이 좋고, 밝은 러셋 색상과 매력적인 외모를 가지고 있습니다. 이 새로운 기술은 또한 실온에서 저장될 수 있는 편리한 물고기 간식 음식을 제공하기 위하여 그밖 물고기의 가공에 적용될 수 있었습니다. 그것은 해양 및 민물고기 모두에 적합합니다.

적당한 소금 농도, 절인 시간과 온도는 제품의 질감을 더 좋게 만듭니다. 가장 좋은 절임 조건은 다음과 같습니다 : 양념 용액에서 8 %의 염 농도; 2 시간의 양념 시간; 10 °C의 절임 온도. 그림1을 참조하십시오.
건조 공정에서 최종 수분 함량과 건조 온도는 질감과 감각 품질에 영향을 줄 수 있습니다. 최상의 건조 조건은 다음과 같습니다: 55%의 건조된 생?...

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

Preparation of High-Quality Fermented Fish Product

이 프로토콜의 목적은 사카로미세세스 세레비시아의접종을 기반으로 한 산업화된 어류 발효 기술을 제공하는 것입니다.

고품질 발효 생선 제품 준비

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

이 절차는 와인 향을 가진 발효 생선 제품의 고품질을 생산하고, 산업 생산에 적용 될 수있다. 이 기술은 생선 제품에 매력적인 외관, 독특한 맛, 맛있는 맛, 좋은 질감 특성을 제공합니다. 그리고 이 제품은 실온에서 1년 이상 보관할 수 있습니다.이 절차를 시작하려면 텍스트 프로토콜에 설명된 대로 해동, 깨끗하고 피부냉동 철갑상어 고기. 물고기를 큐브로 자른다. 다음으로 생선 큐브를 6~12%의 소금 용액과 1대1의 비율로 1~3시간 동안 1대1로 섞는다.그런 다음 양념된 생선 큐브를 스테인레스 스틸 격자 판에 넣고 6~10시간 동안 섭씨 40~60도 사이의 온도에서 뜨거운 공기로 건조시다. 향신료 액체를 준비하려면 1.2 단위의 파를 포함하여 향신료를 섞고, 생강 1 단위, 스타 아니스 0.6 단위, 회향 0.6 단위, 녹차 0.3 단위, 물 40 단위의 후추 0.3 단위를 혼합하십시오. 반 시간 동안 끓여서 여과물을 수집합니다.맛 용액을 준비하려면 향신료 액체, 노란 와인, 화이트 와인, 지팡이 설탕, 글루타민산 나트륨 및 식초를 여기에 표시된 대량 분획에 섞습니다. 그 후, 말린 생선 큐브를 향료 용액과 섞어 0.4~1.6%의 사카로미세스 세레비시아를 일대일 비율로 혼합한다. 용기에 혼합물을 밀봉하고, 6 ~10 시간 동안 섭씨 25 ~35도 사이의 온도에서 발효하십시오.발효 된 생선 큐브를 양념 및 발효 솔루션으로 진공 패키지에 넣고 진공 포장을 0.085 메가 파스칼의 진공 강도, 3.6 초의 밀봉 시간 및 5.5 초의 냉각 시간으로 밀봉하십시오. 그런 다음 10~20분 동안 섭씨 115도에서 121도 사이의 살균 온도를 사용하여 포장된 생선 큐브를 살균합니다. 이 연구에서는 고품질 발효 생선 제품이 준비됩니다.적절한 소금 농도, 양념 시간 및 온도를 사용하면 제품 텍스처 품질이 향상됩니다. 가장 좋은 양념 조건은 8 %의 소금 농도로 보였고 양념 용액, 2 시간의 양념 시간. 그리고 섭씨 10도의 양념 온도.건조 공정에서 최종 수분 함량과 건조 온도는 질감과 센서 품질에 영향을 미칠 수 있습니다. 가장 좋은 건조 조건은 55 %의 건조 된 생선 큐브의 수분 함량과 섭씨 50도의 건조 온도인 것 같습니다. 발효 과정에서, 사카로미세스 세레비시아, 온도 및 시간의 첨가는 생선의 맛, 맛, 색상 및 질감특성에 영향을 미칩니다.성공적인 발효 생선은 맛, 맛, 색상 및 질감에 대한 높은 점수를 가지고 있으며, 그 결과 조화로운 맛, 발효 향료, 와이너리, 러셋 색상, 밝은 표면, 캐러멜 및 알 덴테 질감이 있습니다. 실패한 생선 제품은 맛, 맛, 색상 및 질감에 대한 낮은 점수를 가지고 있어 맛이 좋지 않은 맛, 쓴 맛, 신맛, 비린내, 창백한 색상, 거친 표면, 거친 입맛 및 질감이 악화됩니다. 적당한 살균 및 강도는 4.5 분 인 것처럼 보였습니다.그리고 적당한 살균 온도는 섭씨 121도인 것 같았다. 그런 다음 저장소 고려 사항이 계산되고 텍스트 프로토콜에 설명된 대로 결정됩니다. 그리고 유통 기한에 대한 예측은 섭씨 25도에서 264 일 또는 섭씨 35도에서 205 일입니다.이 절차를 수행할 때 발효 및 살균 단계 모두이 기술의 성공에 필수적이라는 점에 유의해야 합니다. 이 절차를 학습 한 후, 그것은 다른 물고기 종의 발효에 사용할 수 있습니다. 미래에, 그밖 긴장으로 발효는 조사될 수 있었습니다.유산균, 황색포도상구균, 등 세테라와 같은. 지역은 매우 높은 온도로 인해 해를 끼칠 수 있으므로 생선 제품을 살균 할 때 예방 조치를 취해야합니다.

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.

양적 응용 프로그램에 대한 균일 MALDI 시료의 제조

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

연구

생화학

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.

Chromatin Immunoprecipitation을위한 Cross-linked Skeletal Musculus로부터 고품질 핵을 준비하기위한 Filtration 기반 방법

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.

플라바놀에서 플라보놀의 생합성은 원팟 비엔자이매틱 캐스케이드를 확립하여

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.

유전자 발현 프로파일링을 위한 수정된 방법으로 시리얼 씨앗에서 고품질 전사 데이터 획득

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.

단일 분자 물고기에 의한 전사 활성 알레일의 단일 세포 분석

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.

단편 기반 리드 디스커버리에서 스크리닝을 위한 나노 차분 스캔 형광

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-EM용 고온 시료 그리드 준비

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.

글리코겐 구조 측정을 위한 간 글리코겐 분자 추출

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.

미토콘드리아 DNA 합성을 위한 siRNA 혼합물의 제조

In Situ Cellular Cryotomography를 위한 Electron Microscopy Grids 준비

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.

저자 스포트라이트: 향상된 초저온 전자 단층촬영을 위한 그리드 준비 최적화 

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).

포유류 세포의 In Situ Cryotomography를 위한 시료 전처리

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