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

Denne forskningen ble økonomisk støttet av øremerket fondet for Jiangsu Natural Science Fund (BK20170185), prosjekt fra Jiangsu Fisheries administrator (Y2017-30), National Natural Science Foundation i Kina (NFSC31801575), Kina agricultures Research System (CARS-45-26), nasjonale førsteklasses disiplin program av Food Science and Technology (JUFSTR20180201), og Yi Tong-Jiangsu postdoktor program.

1. prøve forberedelser
<ol><li>Tin frossen Sturgeon kjøtt under 20 ° c. Deretter ren, hud av og skjær fisken i terninger (2 cm × 2 cm × 1,5 cm). Merk: råvare kan være fersk eller frossen Sturgeon kjøtt.</li>
	<li>Bland fisk kuber med 6-12% (w/v) saltløsning med en hastighet på 1:1 (fisk kube masse til løsnings volum), ved 10 ° c for 1-3 h.</li>
	<li>Sett marinert fisken kuber på en rustfri stål gridding plate og tørrfisk kuber ved 40-60 ° c i ca 6-10 h med varm luft, eller tørk fisken kuber i vakuu...

<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>

I denne studien, en ny teknikk for produksjon av høykvalitets gjæret fisk produkt med vin aroma og tester for sensoriske egenskaper er gitt. De viktigste prosessene i denne teknikken er marinering, tørking, gjæring og sterilisering. I marinering prosessen påvirker salt konsentrasjonen, temperaturen og tiden alle de textural egenskapene til fisken. Hardheten og chewiness av produktet øker gradvis med økningen av saltkonsentrasjon (0-12%, w/v) og forlengelse av marinering tid (0-2.5 h); men økningen etter 2 h er ik...

Nåværende kommersielle marinert fisk produkt i Kina har problemet med tung salt smak, utilstrekkelig vin aroma, dårlig elastisitet og blek farge, noe som reduserer aksept til forbrukerne. Derfor, en ny teknikk for en høy-kvalitet fisken kjøtt fabrikat med vin aroma nødvendig å bli skapt og optimert.
I de senere årene, har anvendelse av moderne gjæring teknikker i kjøtt og fisk tiltrukket oppmerksomhet fra flere og flere forskere1,2,3,4. Ved inoculation av Start kulturer til kjøtt og fisk, har mattrygghet blitt forbedret, behandlingstiden er forkortet; og produktets sensoriske egenskaper er endret. Saithong et al.5 isolert Lactobacillus bakterier (Lab) fra naturlig plaa-som og brukt denne Lab som en for rett kultur, som indusert høy Surhet og undertrykt patogene bakterier. Zeng et al.6 rapporterte at inoculation med autochtonous for rett kulturer reduserte gjærings tiden og forbedret sanse egenskapene til prøvene. Casaburi et al.7 hevdet at bruk av mikrobielle starter kulturer påvirker utviklingen av aroma i gjæret kjøtt. I disse starter kulturer, S. cerevisiae kunne produsere vin aroma av alkoholholdige gjæring og kan også gi produktet andre forbedrede sensoriske kvaliteter. Derfor er s. cerevisiae en passende startkultur for vin-aroma produkter8,9,10 og vin-aroma fisk produktet kan gjøres av S. cerevisiae.
I prosessen med å lage vin-aroma fisk produkt, teksturen av kjøtt og fisk kan bli påvirket av saltinnhold, vanninnhold, pH, protein denaturering, etc. Derfor kan marinering, tørking, gjæring og sterilisering alle påvirke tekstur egenskaper. Dannelsen av smaken og smak er komplisert og er i hovedsak påvirket av marinering og gjæring, fordi det er svært relatert til hydrolyse av karbohydrater, proteiner og lipider, og milde lipid oksidasjon11,12. Det kan også bli påvirket av tilsetning av krydder13. For utvikling av farge, oppstår Maillard reaksjon som er involvert i prosessen med gjæring og sterilisering10.
Denne artikkelen kan gi teknisk støtte for industrialisering av gjæret fisk produkt med vin aroma, som er av stor betydning for utviklingen av fisken prosessering industrien. Denne teknikken kan forbedre smaken av produktet ved økt proteinolyse (flere frie aminosyrer og TCA-løselig peptider), endre smaken hovedsakelig av alkoholer (etanol, 1-octen-3-OL, 2-metyl-1-propanol og 3-metyl-1-butanol), estere (etanol acetate) og aldehyder (nonanal, 3-methylbutanal og benzaldehyde), øke mouthfeel av høyere hardhet, spenst, gumminess og chewiness, og gi mer attraktiv russet farge av og en lys overflate14. Det gir også forbrukerne bekvemmelighet fordi produktet kan lagres ved romtemperatur. Som beskrevet av andre tidligere studier15,16,17, gjæring med S. cerevisiae har også vist seg å betydelig forbedre sensoriske kvaliteter i andre kjøtt eller fiskeprodukter.
Det er verdt å merke seg at den innførte protokollen kan også brukes i andre arter av fisk, for eksempel gress karpe, sølv karpe, svart karpe, bighead karpe, torsk, laks, etc. For høy kvalitet på fiskeprodukter, bør fisk uten behandling brukes, for eksempel fersk fisk, fisk i is eller frossen fisk lagret i mindre enn 1 år. Dessuten, siden mild lipid oksidasjon kunne forbedre smaken mens omfattende lipid oksidasjon bringer ubehagelig smak, er fisk med mindre fett foretrukket eller skummet fisk anbefales.

<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

Denne protokollen gir en metode for utarbeidelse av industrialiserte gjæret fisk produkt med Sturgeon (Aquilaria sinensis) kjøtt produkt. Prosedyrene var: (1) forbehandling av oppdretts Sturgeon inkludert halshogging, evisceration, skinning-off, rengjøring og skjæring; (2) marinering fisken terningene inne 6-12% (w/v) saltløsning (1:1, fisken terningen masse å løsning kvantum); (3) tørking av fisken kuber til et vanninnhold på 50-60% av varm luft (40-60 ° c) eller ved vakuum; (4) gjæring som involverer vaksinere fisk kuber med 0,4-1,6% (w/w) S. cerevisiae i smaken løsning til fisk kuber og gjæring ved 25-35 ° c for 6-10 h; (5) tetting av fiske kuber i vakuum pakker med marinering og gjærings løsninger; (6) sterilisering ved 115-121 ° c for 10-20 min. Den Sturgeon kjøtt produktet utarbeidet av denne metoden har deilig smak som er mellow og tykk, har ulike typer og store mengder flyktige smaken forbindelser som alkoholer og estere som kan maskere muggen og ubehagelig lukt fra fisk, har moderat salt innhold, men god tekstur egenskaper som høy spenst, gumminess og chewiness, og har lyse russet farge og attraktivt utseende. Denne ny teknikk kunne likeledes være anvendt inne behandlingen av annet fisken å skaffe bekvem fisken snack matvarerne hvilke det kan være lagret for romtemperatur. Det er hensiktsmessig for både marine og ferskvannsfisk.

Den egnede salt konsentrasjonen, marinering tiden og temperaturen gjør produktets textural kvalitet bedre. De beste marinering forholdene var som følger: salt konsentrasjonen av 8% i den marinering løsningen; marinering tid på 2 timer; og marinering temperatur på 10 ° c. Se figur 1.
I tørkeprosessen kan det endelige Fuktinnholdet og tørke temperaturen påvirke teksturen og sanse kvaliteten. De beste tørke forholdene var som følger: fuktinnhold av tørket...

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

Preparation of High-Quality Fermented Fish Product

Målet med protokollen er å gi en industrialiserte fiske gjæring teknikk basert på inoculation av Saccharomyces cerevisiae.

Utarbeidelse av høy kvalitet gjæret fisk produkt

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

Denne prosedyren produserer en høy kvalitet av gjæret fiskeprodukt med vin aroma, og kan brukes i industrien produksjon. Denne teknikken gir fiskeproduktet et attraktivt utseende, unik smak, deilig smak og gode teksturegenskaper. Og dette produktet kan lagres ved romtemperatur i minst ett år.For å begynne denne prosedyren, tine, ren og hud frosset stør kjøtt som beskrevet i tekstprotokollen. Skjær fisken i terninger. Bland deretter fiskebitene med seks til 12% saltløsning i forholdet mellom en til en ved 10 grader Celsius i en til tre timer.Deretter legger du de marinerte fiskebitene på en risteplate i rustfritt stål og tørker dem med varm luft ved en temperatur mellom 40 og 60 grader Celsius i seks til 10 timer. For å forberede kryddervæsken, bland krydder, inkludert 1,2 enheter av scallions, en enhet ingefær, 0,6 enheter stjerne anis, 0,6 enheter fennikel, 0,3 enheter grønn te og 0,3 enheter pepper med 40 enheter vann. Kok i en halv time, og samle filtrat.For å forberede smaksløsningen, bland kryddervæske, gulvin, hvitvin, sukkerrør, mononatriumglutamat og eddik i massefraksjonene som vises her. Etter dette blander du de tørkede fiskebitene med smaksløsningen, inokulerer 0,4 til 1,6% saccharomyces cerevisiae i forholdet mellom en-til-en. Forsegle blandingen i en beholder, og gjær ved en temperatur mellom 25 og 35 grader Celsius, i seks til 10 timer.Legg de fermenterte fiskebitene i en vakuumpakke med marinerings- og gjæringsløsninger og forsegle vakuumpakken med en vakuumstyrke på 0,085 megapascals, en tetningstid på 3,6 sekunder og en kjøletid på 5,5 sekunder. Steriliser deretter de pakkede fiskebitene ved hjelp av en steriliseringstemperatur mellom 115 og 121 grader Celsius i 10 til 20 minutter. I denne studien er gjæret fiskeprodukt av høy kvalitet tilberedt.Ved hjelp av en passende saltkonsentrasjon, marineringstid og temperatur gjør produktene teksturkvalitet bedre. De beste marineringsforholdene ser ut til å være en saltkonsentrasjon på 8%Og marineringsløsningen, en marineringstid på to timer. Og en marinering temperatur på 10 grader Celsius.I tørkeprosessen kan det endelige fuktighetsinnholdet og tørketemperaturen påvirke teksturen og sensorkvaliteten. De beste tørkeforholdene er syntes å være et fuktighetsinnhold i de tørkede fiskebitene på 55%og tørketemperatur på 50 grader Celsius. I gjæringsprosessen, tilsetning av saccharomyces cerevisiae, temperaturen, og tiden alle effekt smak, smak, farge, og tekstur egenskaper av fisk.Den vellykkede fermenterte fisken har høye score for smak, smak, farge og tekstur, noe som resulterer i harmonisk smak, gjærende duft, vinaktig, russetfarge, lys overflate, karamell og al dente tekstur. Det mislykkede fiskeproduktet har de lave poengsummene for smak, smak, farge og tekstur som resulterer i dårlig smak, bitterhet, surhet, fishiness, blek farge, grov overflate, grov munnfølelse og forverret tekstur. Den passende sterilisering og intensitet er syntes å være 4,5 minutter.Og en passende steriliseringstemperatur ser ut til å være 121 grader Celsius. Lagringshensynene beregnes og bestemmes deretter som beskrevet i tekstprotokollen. Og spådommen for holdbarheten er 264 dager ved 25 grader Celsius eller 205 dager ved 35 grader Celsius.Når du utfører denne prosedyren, er det viktig å merke seg at både gjæring og steriliseringstrinnene er avgjørende for suksessen til denne teknikken. Etter å ha lært denne prosedyren, kan den brukes til gjæring av andre fiskearter. I fremtiden kan gjæring med andre stammer undersøkes.Som lactobacillus, stafylokokker, et cetera. regioner, sørg for å ta forholdsregler ved sterilisering av fiskeprodukter, da ekstremt høy temperatur kan forårsake skade.

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.

Utarbeidelse av homogene MALDI Prøver for Quantitative Applications

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

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.

En filtreringsbasert metode for å fremstille kjernekvalitet av høy kvalitet fra tverrbundet skjelettmuskulatur for kromatinimmunutfelling

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.

Biosyntesen av en Flavonol fra en Flavanone ved å etablere en 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 ...

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.

Skaffe transkripsjonsdata av høy kvalitet fra kornfrø med en modifisert metode for genuttrykksprofilering

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.

Enkeltcelleanalyse av transkripsjonsaktive alleler av enkeltmolekyl fisk

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.

Nano-differensialskanning fluorimetri for screening i fragmentbasert blyoppdagelse

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.

Klargjøring av høytemperatur prøvegitter 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, ...

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.

Ekstraksjonen av leverglykogenmolekyler for bestemmelse av glykogenstruktur

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.

Fremstilling av siRNA-blanding for mitokondriell DNA-syntese

Utarbeidelse av elektronmikroskopigitter for in situ cellulær kryotomografi

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.

Forfatter i søkelyset: Optimalisering av rutenettforberedelse for forbedret kryoelektrontomografi 

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).

Prøvepreparering for in situ kryotomografi av pattedyrceller

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