CN109456922A - The beach the Halophilic Bacterium bacterial strain Hua Jin bacillus of one plant of raising alec fermentation quality - Google Patents
The beach the Halophilic Bacterium bacterial strain Hua Jin bacillus of one plant of raising alec fermentation quality Download PDFInfo
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Abstract
The invention discloses the beach the Halophilic Bacterium bacterial strain Hua Jin bacillus that one plant is improved alec fermentation quality, are related to food microorganisms technical applications.The bacterial strain was deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC) on September 27th, 2018, and number is CGMCC NO.16542, can be described as Bacillus hwajinpoensis CGMCC16542.The bacterial strain utilizes Gibbons Screening of Media from traditional zymotic shrimp paste, which has stronger production protease ability, can be applied to aquatic animal products fermentation, can accelerate the fermentation of aquatic animal products and the improvement of flavor.
Description
Technical field
The present invention relates to the moderate of food microorganisms technical applications more particularly to one plant of raising alec fermentation quality is thermophilic
The beach salt bacteria strain Hua Jin bacillus can effectively facilitate alec fermentation and improve the new strains of alec quality.
Background technique
China's freshwater aquiculture is quickly grown, but is relatively lagged behind in processing industry development.Therefore it develops and is suitble to domestic consumption market
The freshwater fish products of demand have important practical significance to the fresh-water fishes industry sound development of promotion.Currently, in fermentation aquatic product product
Amino acid content is abundant, and nutritive value is high, is loved by consumers, but that there is fermentation times again is long, contains for fermented product
The problems such as salt amount height and unstable fermentation.Halophilic Bacterium is a kind of raw between 0.5mol/L to 2.5mol/L in NaCl concentration
Long optimal extreme microorganism.Compared with slight Halophiles, Halophilic Bacterium can be grown in hypersaline environment, to reduce hair
Living contaminants during ferment.Compared with extremely halophilic archaea, Halophilic Bacterium is wider than the thermophilic salt range of halophilic archaea, is distributed and comes
Source more extensively, to nutriment require also it is lower, be more easily adapted to conform to living environment, in actual production be easier applied.Have
A little Halophilic Bacteriums have stronger extracellular property of protease again.The present invention will utilize one plant of Halophilic Bacterium new strains fermented fish
Meat pulp, it is short to obtain fermentation time, and salinity is lower and flavor and the higher flesh of fish jam products of nutritive value.For aquatic products fermentation quality
Promotion provide theoretical basis and method.
Summary of the invention
The present invention is sieved from traditional zymotic shrimp paste to accelerate the fermentation of aquatic animal products and promote the quality of fermented product
Select the new strains of one plant of improvement fermentation aquatic product product quality.The bacterial strain can effectively improve the flavor and nutriture value of fermentation aquatic product product
Value, provides guidance for the promotion of fermented product quality.
The technical solution used in the present invention is as follows:
Halophilic Bacterium JD (Bacillus hwajinpoensis JD) provided by the invention, to can be applied to ferment
The new strains of alec quality-improving.The Halophilic Bacterium bacterial strain has been deposited in positioned at Beijing's southern exposure on September 27th, 2018
The China Committee for Culture Collection of Microorganisms of the Institute of Microorganism, Academia Sinica of the institute 3 of area North Star West Road 1 is common
Microorganism center (CGMCC), number are CGMCC NO.16542, it is proposed that classification naming: the beach Hua Jin bacillus
hwajinpoensis。
In one aspect of the invention, the use of above-mentioned Bacillus hwajinpoensis CGMCC 16542 is provided
On the way, the promotion for alec ferment local-flavor and quality, it is a kind of to utilize the beach the Halophilic Bacterium bacterial strain Hua Jin fermentation of bacillus flesh of fish
The method of sauce carries out as steps described below:
(1) processing of raw material: minced fish is chopped off using cutmixer, marinated sea salt 5~20% (w/w) is added, before fermentation
Be placed on 5 in 4 DEG C of refrigerators~for 24 hours.
(2) preparation of Halophilic Bacterium bacterial strain: by Halophilic Bacterium Bacillus hwajinpoensis CGMCC
16542 are seeded in activation culture on culture medium, and repetitive operation is three times.The strain bacterium solution of culture is centrifuged in 4 DEG C, 10000r/min
Then 10min is washed twice with sterile saline, be resuspended in sterile saline.Finally bacterial concentration is adjusted to
105~106CFU/mL saves bacterium solution in 4 DEG C, and uses in interior for 24 hours.
(3) addition of leavening: Halophilic Bacterium strain Bacillus hwajinpoensis CGMCC 16542 is pressed
Final additive amount (104~107CFU/g it) is mixed into pretreated raw material fish, so that final bacterium number is reached addition and require
(4) fish meat paste is fermented: the mixed material that step (3) is obtained 5~30d of heat-preservation fermentation at 28~40 DEG C of temperature；
To ferment fish meat paste be placed in it is spare in -20 refrigerators.
(5) stirring in fish meat paste fermentation process and ventilative: in 5~30d of alec fermentation, every 1~5d sterile
Under the conditions of to bagoong carry out 1~5min of clockwise stirring, after stirring stand breathe freely 10~120s.
(6) fish meat paste that step (4) obtains fish meat paste shortening: is steamed into 20~40min to fish using vapor in steaming tray
Meat pulp is well-done.
(7) fish meat paste sterilizes: by the fish meat paste filling and sealing of step (6) shortening, at 120 DEG C two-stage sterilization 15~
20min is to get fish meat paste finished product.
(8) evaluation of product fermentation physical and chemical index: amino acid nitrogen content evaluation, method is referring to GB5009.235-2016
PH meter.TVB-N content evaluation, method is referring to GB5009.228-2016 microdiffusion.The survey of histamine content
Fixed, method is referring to GB5009.208-2016 spectrophotometry.
(8) subjective appreciation: forming subjective appreciation group by 10 people, needs to carry out flavor training to group member before evaluation.It comments
Dividing includes 0~90 scores, is carried out to the fishiness of fish meat paste, fishy smell, fermented flavour, ammonia taste, delicate flavour, saline taste, sweet taste and bitter taste
The comprehensive score of whole flavor.
Beneficial effects of the present invention
Fermentation time using the fish meat paste of the beach Halophilic Bacterium bacterial strain Hua Jin fermentation of bacillus is short, and salinity is lower,
Amino acid nitrogen content is higher, and flavour is delicious, and nutritive value is abundant, and the content of Volatile Base Nitrogen is lower than EU criteria, histamine
Content be far below national standard, be a kind of quality and highly-safe fermented fish meat paste product.
In conclusion the present invention using had been provided with the uniqueness of patent of invention, obtain substantive distinguishing features outstanding and
The innovation and practicality of marked improvement, produces beneficial effect.
Detailed description of the invention
Fig. 1 is Halophilic Bacterium Bacillus hwajinpoensis JD colonial morphology；
Fig. 2 is the systematic growth of the 16SrRNA sequence construct of Halophilic Bacterium Bacillus hwajinpoensis JD
Tree.Note: zzy9 is Halophilic Bacterium Bacillus hwajinpoensis JD of the present invention.
Specific embodiment
1. the separation screening of bacterial strain:
Using the fresh shrimp paste of traditional zymotic as raw material, shrimp paste derives from Tangshan, Hebei Province Fengnan District Hei Yanzi town fisherman
Make shrimp paste, the separation screening Halophilic Bacterium from shrimp paste by oneself.The separation and purifying of Halophilic Bacterium use 1% skimmed milk powder of addition
Gibbons culture medium, the shrimp paste after sterile water is diluted by certain multiple is coated on solid Gibbons culture medium, 30 DEG C
48h is cultivated, picking has the bacterium colony of characteristic feature, continues to carry out on the solid Gibbons culture medium for adding 1% skimmed milk powder
Separating for several times purifying obtains the Halophiles bacterial strain that pure culture produces protease.By separation and strain inoculated after purification is to Gibbons
On inclined-plane, it is spare to be placed in 4 DEG C of refrigerator short term storages by 30 DEG C of culture 48h.
2. the identification of bacterial strain
The morphological feature of 2.1 bacterial strains
Bacterium colony is in crocus, surface is smooth, neat in edge, opaque, compact colonies, and Gram's staining is the positive, thallus
It is in the shape of a rod.After cultivating for 24 hours in liquid Gibbons culture medium, bacterium solution is muddy.
The physiological characteristic of 2.2 bacterial strains
The bacterial strain can be grown when NaCl concentration is 0~20%；When pH is 7~9, growth is very fast；In temperature
Range, which is 25 DEG C~43 DEG C, well to be grown；
2.3 molecular biology identification
The Halophilic Bacterium separated is sequenced through 16SrRNA, gene order through operation arrange after,
It is retrieved in Ezbiocloud, is determined as (Bacillus hwajinpoensis).According to the homologous strain retrieved, application
The Mege5.1 program of DNAStar software constructs biological evolution relational tree such as Fig. 2.
The Halophilic Bacterium bacterial strain has been deposited in positioned at BeiChen West Road, Chaoyang District, BeiJing City 1 on September 27th, 2018
The China Committee for Culture Collection of Microorganisms's common micro-organisms center for the Institute of Microorganism, Academia Sinica that institute 3
(CGMCC), number is CGMCC NO.16542, it is proposed that classification naming: the beach Hua Jin bacillus
hwajinpoensis。
Embodiment 1:
The method of the one plant of beach Halophilic Bacterium bacterial strain Hua Jin fermentation of bacillus fish meat paste carries out as steps described below:
(1) processing of raw material: minced fish is chopped off using cutmixer, and marinated sea salt 5% (w/w) is added, places before fermentation
In 4 DEG C of refrigerators for 24 hours.
(2) preparation of Halophilic Bacterium bacterial strain: by Halophilic Bacterium strain Bacillus hwajinpoensis CGMCC
16542 are seeded in activation culture on culture medium, and repetitive operation is three times.The strain bacterium solution of culture is centrifuged in 4 DEG C, 10000r/min
Then 10min is washed twice with sterile saline, be resuspended in sterile saline.Finally bacterial concentration is adjusted to
105~106CFU/mL saves bacterium solution in 4 DEG C, and uses in interior for 24 hours.
(3) addition of leavening: Halophilic Bacterium strain Bacillus hwajinpoensis CGMCC 16542 is pressed
Final additive amount 104CFU/g it) is mixed into pretreated raw material fish, so that final bacterium number is reached addition and require
(4) fish meat paste is fermented: the mixed material that step (3) is obtained heat-preservation fermentation 30d at 28 DEG C of temperature；It will ferment
Fish meat paste is placed in spare in -20 refrigerators.
(5) stirring in fish meat paste fermentation process and ventilative: in the 30d of alec fermentation, aseptically every 5d
Clockwise stirring 1min is carried out to bagoong, ventilative 10s is stood after stirring.
(6) fish meat paste that step (4) obtains fish meat paste shortening: is steamed into 20min to fish meat paste using vapor in steaming tray
It is well-done.
(7) fish meat paste sterilizes: by the fish meat paste filling and sealing of step (6) shortening, the two-stage sterilization 15min at 120 DEG C, i.e.,
Obtain fish meat paste finished product.
(8) evaluation of product fermentation physical and chemical index: amino acid nitrogen content evaluation, method is referring to GB5009.235-2016
PH meter.TVB-N content evaluation, method is referring to GB5009.228-2016 microdiffusion.The survey of histamine content
Fixed, method is referring to GB5009.208-2016 spectrophotometry.
(9) subjective appreciation: forming subjective appreciation group by 10 people, needs to carry out flavor training to group member before evaluation.It comments
Dividing includes 0~90 scores, is carried out to the fishiness of fish meat paste, fishy smell, fermented flavour, ammonia taste, delicate flavour, saline taste, sweet taste and bitter taste
The comprehensive score of whole flavor.
(10) physical and chemical index of fish meat paste and sensory evaluation
Content using the amino-acid state of this plant of Halophilic Bacterium strain fermentation fish meat paste is 0.639g/100g；Volatility
Alkali nitrogen content is 24.87mg/100g, is lower than EU criteria 35mg/100g；Histamine content is 3.82mg/100g, far below food
The content 20mg/100g of histamine in the safe national standard fish of product；The comprehensive score of sensory evaluation is 6.33.It is not added with bacterial strain fish
The sensory evaluation scores of sauce are 5.67.Bilgy odour intrinsic in raw material greatly reduces in this fish meat paste, and delicate flavour is obvious, and is not added with bacterial strain
Alec is compared, and flavor is obviously improved.
Embodiment 2:
The method of the one plant of beach Halophilic Bacterium bacterial strain Hua Jin fermentation of bacillus fish meat paste carries out as steps described below:
(1) processing of raw material: minced fish is chopped off using cutmixer, and marinated sea salt 20% (w/w) is added, places before fermentation
In 4 DEG C of refrigerators for 24 hours.
(2) preparation of Halophilic Bacterium bacterial strain: by Halophilic Bacterium strain Bacillus hwajinpoensis CGMCC
16542 are seeded in activation culture on culture medium, and repetitive operation is three times.The strain bacterium solution of culture is centrifuged in 4 DEG C, 10000r/min
Then 10min is washed twice with sterile saline, be resuspended in sterile saline.Finally bacterial concentration is adjusted to
105~106CFU/mL saves bacterium solution in 4 DEG C, and uses in interior for 24 hours.
(3) addition of leavening: Halophilic Bacterium strain Bacillus hwajinpoensis CGMCC 16542 is pressed
Final additive amount 107CFU/g it) is mixed into pretreated raw material fish, so that final bacterium number is reached addition and require
(4) fish meat paste is fermented: the mixed material that step (3) is obtained heat-preservation fermentation 5d at 40 DEG C of temperature；It will ferment
Fish meat paste is placed in spare in -20 refrigerators.
(5) stirring in fish meat paste fermentation process and ventilative: aseptically right every 1d in the 5d of alec fermentation
Bagoong carries out clockwise stirring 5min, and ventilative 120s is stood after stirring.
(6) fish meat paste that step (4) obtains fish meat paste shortening: is steamed into 40min to fish meat paste using vapor in steaming tray
It is well-done.
(7) fish meat paste sterilizes: by the fish meat paste filling and sealing of step (6) shortening, the two-stage sterilization 25min at 120 DEG C, i.e.,
Obtain fish meat paste finished product.
(8) evaluation of product fermentation physical and chemical index: amino acid nitrogen content evaluation, method is referring to GB5009.235-2016
PH meter.TVB-N content evaluation, method is referring to GB5009.228-2016 microdiffusion.The survey of histamine content
Fixed, method is referring to GB5009.208-2016 spectrophotometry.
(9) subjective appreciation: forming subjective appreciation group by 10 people, needs to carry out flavor training to group member before evaluation.It comments
Dividing includes 0~90 scores, is carried out to the fishiness of fish meat paste, fishy smell, fermented flavour, ammonia taste, delicate flavour, saline taste, sweet taste and bitter taste
The comprehensive score of whole flavor.
(10) physical and chemical index of fish meat paste and sensory evaluation
Content using the amino-acid state of this plant of Halophilic Bacterium strain fermentation fish meat paste is 0.312g/100g；Volatility
Alkali nitrogen content is 7.80mg/100g, is lower than EU criteria 35mg/100g；Histamine content is 1.98mg/100g, far below food
The content 20mg/100g of histamine in the safe national standard fish of product；The comprehensive score of sensory evaluation is 5.93.It is not added with bacterial strain fish
The sensory evaluation scores of sauce are 5.48.Bilgy odour intrinsic in raw material greatly reduces in this fish meat paste, and delicate flavour is obvious, and is not added with bacterial strain
Alec is compared, and flavor is obviously improved.
Claims (2)
1. Halophilic Bacterium bacterial strain, deposit number is CGMCC NO.16542, it is proposed that classification naming: the beach Hua Jin bacillus
Bacillus hwajinpoensis。
2. a kind of using described in claim 1 a kind of using the beach Halophilic Bacterium bacterial strain Hua Jin fermentation of bacillus fish meat paste
Method, it is characterised in that carry out as steps described below:
(1) processing of raw material: minced fish is chopped off using cutmixer, and marinated sea salt 5~20% (w/w) is added, places before fermentation
5 in 4 DEG C of refrigerators~for 24 hours；
(2) preparation of Halophilic Bacterium bacterial strain: Halophilic Bacterium Bacillus hwajinpoensis CGMCC 16542 is connect
Kind activation culture on culture medium, repetitive operation is three times；The strain bacterium solution of culture is centrifuged 10min in 4 DEG C, 10000r/min,
Then it is washed twice, is resuspended in sterile saline with sterile saline；Bacterial concentration is finally adjusted to 105~
106CFU/mL saves bacterium solution in 4 DEG C, and uses in interior for 24 hours；
(3) addition of leavening: by Halophilic Bacterium strain Bacillus hwajinpoensis CGMCC 16542 by final
Additive amount (104~107CFU/g it) is mixed into pretreated raw material fish, so that final bacterium number is reached addition and require
(4) fish meat paste is fermented: the mixed material that step (3) is obtained 5~30d of heat-preservation fermentation at 28~40 DEG C of temperature；It will hair
The good fish meat paste of ferment is placed in spare in -20 refrigerators；
(5) stirring in fish meat paste fermentation process and ventilative: in 5~30d of alec fermentation, every 1~5d in aseptic condition
Under to bagoong carry out 1~5min of clockwise stirring, after stirring stand breathe freely 10~120s；
(6) fish meat paste that step (4) obtains fish meat paste shortening: is steamed into 20~40min to fish meat paste using vapor in steaming tray
It is well-done；
(7) fish meat paste sterilizes: by the fish meat paste filling and sealing of step (6) shortening, 15~20min of two-stage sterilization at 120 DEG C, i.e.,
Obtain fish meat paste finished product.
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| CN201811452816.2A CN109456922B (en) | 2018-11-30 | 2018-11-30 | Moderately halophilic bacteria strain Zhan beach bacillus for improving fermentation quality of fish paste |
| US16/761,132 US11974590B2 (en) | 2018-11-30 | 2019-03-29 | Method of fermentation of fish paste by medium halophilic bacteria |
| PCT/CN2019/080282 WO2020107760A1 (en) | 2018-11-30 | 2019-03-29 | Moderately halophilic bacteria and method for fermenting fish meat sauce by using same |
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| CN201811452816.2A CN109456922B (en) | 2018-11-30 | 2018-11-30 | Moderately halophilic bacteria strain Zhan beach bacillus for improving fermentation quality of fish paste |
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| CN111534451A (en) * | 2020-01-13 | 2020-08-14 | 江苏大学 | Planococcus fermentation agent for improving fermentation quality of low-salt fish gravy |
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