阅读说明：本技术 一种臭氧联合隔氧微生物发酵的贝肉生物脱腥方法 (Method for biologically removing fishy smell of shellfish meat through ozone combined with anaerobic microbial fermentation ) 是由 何志刚 林晓姿 梁璋成 苏昊 李维新 庄林歆 于 2019-08-30 设计创作，主要内容包括：本发明公开了一种臭氧联合隔氧微生物发酵的贝肉生物脱腥方法,主要步骤为冻藏或新鲜贝肉,经酸性饱和臭氧水减菌处理,添加糖水作为碳源补充,接种具有生物脱腥效果的副干酪乳杆菌副干酪亚种RP38和酿酒酵母JJ4菌株,采取充二氧化碳或充氮隔氧的方式进行厌氧生物发酵脱腥。本发明规避了贝肉在生物脱腥过程的杂菌污染腐败风险,确保优势脱腥菌种的生物发酵顺利进行,获得带有清淡发酵果香、组织结构完整、没有明显腥味的脱腥贝肉。(The invention discloses a method for biologically removing fishy smell of shellfish meat through ozone combined anaerobic microbial fermentation, which mainly comprises the steps of freezing or storing fresh shellfish meat, carrying out bacteria reduction treatment by acidic saturated ozone water, adding sugar water as a carbon source for supplement, inoculating lactobacillus paracasei subspecies paracasei RP38 and saccharomyces cerevisiae JJ4 bacterial strains with biological fishy smell removal effect, and carrying out anaerobic biological fermentation and fishy smell removal by adopting a carbon dioxide or nitrogen and oxygen filling mode. The method avoids the risk of mixed bacteria pollution and putrefaction of the shellfish meat in the biological fishy smell removing process, ensures that the biological fermentation of the dominant fishy smell removing strain is smoothly carried out, and obtains the fishy smell removed shellfish meat with light fermentation fruity flavor, complete tissue structure and no obvious fishy smell.)

1. A method for biologically removing fishy smell of shellfish meat fermented by ozone combined with an oxygen-insulated microorganism is characterized by comprising the following steps: which comprises the following steps:

1) the shellfish meat is subjected to ozone sterilization treatment to reduce the bacterial contamination base number of the raw materials;

2) adding fermentation liquor into the treated shellfish meat, inoculating lactobacillus and Saccharomyces cerevisiae with biological fishy smell removing effect as leaven, and performing anaerobic microbial fermentation by charging carbon dioxide or nitrogen and oxygen.

2. The method for biologically deodorizing the shellfish meat fermented by the ozone combined with the anaerobic microorganism according to claim 1, characterized by comprising the following steps: the shellfish meat includes fresh meat of scallop, mussel and clam and frozen meat thereof.

3. The method for biologically removing the fishy smell of the shellfish meat fermented by the ozone combined with the oxygen-insulated microorganism according to claim 1, wherein the step 1) comprises adding saturated ozone water into the shellfish meat according to the mass ratio of 1:1.0 ~ 1.5.5, adjusting the pH value of the ozone water to 4.5 ~ 4.6.6 by using citric acid, adjusting the temperature to 5-10 ℃ by using ice blocks, treating for 20 ~ 25min, taking out the shellfish meat, and draining.

4. The method for biologically removing the fishy smell of the shellfish meat fermented by the ozone combined with the anaerobic microorganism according to claim 1, wherein the fermentation liquid in the step 2) is prepared by preparing sugar water with a mass concentration of 1 ~ 2% by using white granulated sugar, adjusting the pH to 4.5 ~ 4.6.6 by using citric acid, sterilizing and cooling for later use.

5. The method for biologically deodorizing the shellfish meat fermented by the ozone combined with the anaerobic microorganism according to claim 1, characterized by comprising the following steps: step 2) the lactobacillus is lactobacillus paracasei subspecies paracasei RP38 (with facultative anaerobic biological deodorization capacity)Lactobacillus paracasei subsp. paracasei RP38) which is preserved in China center for type culture Collection with the preservation number of CCTCC NO of M2019322 and the preservation date of 2019, 5 months and 5 days;

the saccharomyces cerevisiae is saccharomyces cerevisiae JJ4 with fragrance producing and fishy smell removing capabilities, is preserved in China center for type culture collection, and has a preservation number of CCTCC NO: m2018219, preservation date of 2018, 4 months and 19 days.

6. The method for biologically deodorizing shellfish meat fermented by ozone combined with an oxygen-insulated microorganism according to claim 5, wherein the method comprises the following steps: the preparation steps of the leaven in the step 2) are as follows: activating Lactobacillus paracasei subspecies paracasei RP38, inoculating in MRS liquid culture medium or molasses culture medium, and culturing at 25-30 deg.C until the bacterial count reaches 1.0 × 10⁹More than cfu/mL, centrifuging to obtain fresh lactic acid bacteria, and directly using as a leavening agent or preparing into a direct-vat-set microbial inoculum for use;

activating Saccharomyces cerevisiae JJ4, inoculating to molasses culture medium, and culturing at 25-30 deg.C until bacterial count reaches 1.0 × 10⁸More than cfu/mL, then centrifuging to obtain fresh yeast which is directly used as a leavening agent or prepared into a direct-vat set microbial inoculum for use.

7. The method for biologically deodorizing shellfish meat through ozone combined with anaerobic microorganism fermentation according to claim 6, characterized in that the centrifugation is carried out at 4500 ~ 5000r/min for 5-8 min.

8. The method for biologically removing the fishy smell of the shellfish meat through the ozone-combined oxygen-insulated microorganism fermentation according to claim 5, wherein the step 2) comprises the steps of containing the shellfish meat treated in the step 1) and the fermentation liquid in a biological fermentor which can be hermetically sealed, inoculating the composite bacteria containing lactobacillus paracasei subspecies paracasei RP38 and saccharomyces cerevisiae JJ4 with the inoculation amount of 1% ~ 5% (v/w), and filling the fermentor with CO after the inoculation₂Or N₂Sealing in oxygen-isolated manner, and performing anaerobic microbial fermentation at 25 ~ 30 deg.C for 3 ~ 6h to obtain deodorized shellfish meat.

9. The method for biologically deodorizing shellfish meat fermented by ozone combined with oxygen-insulated microorganisms according to claim 8, wherein the mass ratio of the shellfish meat to the fermentation broth is 1:0.5 ~ 1.

10. The method for biologically removing the fishy smell of the shellfish meat through the ozone combined with the anaerobic microorganism fermentation is characterized in that the ratio of lactobacillus paracasei subspecies paracasei RP38 to saccharomyces cerevisiae JJ4 in the composite bacteria is 3 ~ 5: 1.

Technical Field

The invention belongs to the field of aquatic product processing, and particularly relates to a method for biologically removing fishy smell of shellfish meat through ozone combined with anaerobic microorganism fermentation.

Background

Aquatic products have certain unpleasant volatile smells such as fishy smell, earthy smell and the like to different degrees, and the application of shellfish in the processing of leisure food is greatly restricted in aquatic products to a great extent, for example, scallop is a marine product processed and dried by adductor muscle of scallop, and the scallop is white, slightly yellow in middle, shaped like a cylinder, elastic, extremely sweet in taste and rich in protein, lipid, iodine, iron and other mineral substances; the hard clam is also called as veneriforms philippinarum, yellow clam, sea clam and the like, is distributed in the south and north coastal areas of China, contains high protein and low fat in meat, is rich in nutrition, delicious and tasty, contains rich taurine, has good nutrition and health care functions, and is deeply loved by consumers at home and abroad; mussels are commonly called as rainbow and mussel, are high-quality marine cultured shellfish, have high edible value, are rich in protein, have 8 essential amino acids such as valine and leucine required by human bodies, have amino acid compositions very similar to those of human bodies, and have multiple health-care functions. Shellfish aquatic products are mostly fresh foods, some shellfish aquatic products are made into dry products or cans, the palatability of fresh and dried shellfish meat is poor, the fishy smell is heavy, the acceptability of inland people is low, and the limitation of sales areas is caused. Propionaldehyde, pentanal, octanal, nonanal and other aldehydes are considered as main fishy smell components of aquatic products, C4-C7 aldehydes are mainly from fat oxidative degradation, threshold values of the aldehydes are lower than those of other compounds, and when the threshold values are higher than the threshold values, pungent smell or fishy smell is presented, and the octanal and the nonanal are proved to be products of oleic acid oxidation, and when the content is high, pungent smell is presented.

The fishy smell removing method of the aquatic products mainly comprises a chemical method, a physical method, a biological method, a masking method and the like, wherein one method is used independently, the fishy smell removing effect is not obvious, and two or more mixed fishy smell removing technologies are mostly adopted. Research reports that the anti-oxidation effect of theanine, theaflavin and thearubigin on free fatty acid in the shellfish meat can reduce fishy smell components; the ginger juice can dissolve aldehyde substances, reduce fishy smell, cover up and remove fishy smell of aquatic products, but has pungent taste; performing Angel yeast fermentation biological deodorization on the saury soup, and reducing the content of aldehydes such as nonenal, E-2, 6-nonenal, E-2-nonenal, 2, 4-nonenal, E-2-decenal, E-2, 4-decadienal, E-2, 4-undecadinaldehyde, E-2, 4-octenal, dodecanal, 2-dodecenal and the like, dienal and the like in the fish soup so as to weaken the fishy smell of the fish soup; ozone is a strong oxidant, can be degraded to generate extremely active monatomic oxygen and hydroxyl free radicals with strong oxidation effect, can oxidize and decompose terpenes, aldehydes, alcohols, amines, pyridines and other substances, and can be converted into substances without fishy smell or with a high fishy smell threshold value.

Disclosure of Invention

The aquatic fish and shellfish protein and other nutrient substances are rich, and are easy to be polluted by putrefying microorganisms to generate putrefying substances in the processing process, therefore, when the biological fishy smell removal of the shellfish meat is carried out, a microbial fermentation biological fishy smell removal process is needed, the growth of putrefying bacteria is inhibited, and the fishy smell removal effect is achieved, so that the risk of deterioration caused by the growth of the putrefying bacteria in the biological fishy smell removal process of the shellfish meat is solved, and the rapid propagation of target microorganisms and the fishy smell removal effect can be ensured. In view of the above, the present invention aims to provide a method for biologically deodorizing shellfish meat through ozone combined with anaerobic microorganism fermentation.

In order to achieve the purpose, the invention adopts the following technical scheme;

a method for biologically removing fishy smell of shellfish meat by ozone combined with anaerobic microorganism fermentation comprises the following steps: 1) carrying out bacteria reduction treatment on the shellfish meat by using acidic saturated ozone water to ensure that the mixed bacteria base number of the raw materials is reduced by using the maximum ozone concentration; 2) adding carbon source fermentation liquor into the shellfish meat, and inoculating lactobacillus and saccharomyces cerevisiae with biological deodorization effect as leavening agents; 3) anaerobic microorganism fermentation is carried out by adopting a carbon dioxide filling or nitrogen filling and oxygen isolation mode to inhibit the proliferation of aerobic bacteria.

The shellfish meat includes fresh meat of scallop, mussel and clam and frozen meat thereof.

The specific steps of the ozone sterilization treatment are as follows: adding saturated ozone water into the shellfish meat according to the mass ratio of 1: 1.0-1.5, adjusting the pH value of the ozone water to 4.5-4.6 by using citric acid, adjusting the temperature to 5-10 ℃ by using ice blocks, treating for 20-25 min, fishing out the shellfish meat, and draining.

The microorganism used in the biological fermentation deodorization process is the lactobacillus paracasei subspecies paracasei RP38 which is selected and preserved independently, is facultative anaerobic and has the biological deodorization capacity, and has the functions of producing fragrance and deodorizingSaccharomyces cerevisiae JJ4 with fishy smell removing capability. The lactobacillus paracasei subspecies paracasei RP38 is preserved in China Center for Type Culture Collection (CCTCC) with the preservation number of M2019322 and the preservation date of 2019, 5 months and 5 days; saccharomyces cerevisiae JJ4, which is preserved in China center for type culture Collection with the preservation number of CCTCC NO: m2018219, preservation date of 2018, 4 months and 19 days. The preparation steps of the leaven are as follows: activating subspecies RP38 of lactobacillus paracasei and paracasei, inoculating the subspecies RP38 in MRS liquid culture medium or molasses culture medium, and culturing at 25-30 deg.C until the bacterial quantity reaches 1.0 × 10⁹More than cfu/mL, centrifuging for 5-8min at the rotating speed of 4500-5000r/min to obtain fresh lactic acid bacteria, and directly using as a leavening agent or preparing into a direct-vat-set microbial inoculum; ② after activating the saccharomyces cerevisiae JJ4, inoculating in molasses culture medium, culturing at 25-30 ℃ until the bacterial quantity reaches 1.0 multiplied by 10⁸More than cfu/mL, and then centrifuging for 5-8min at the rotating speed of 4500-5000r/min to obtain the fresh yeast which is directly used as a leavening agent or prepared into a direct-vat set microbial inoculum for use.

The shellfish protein and other nutrient substances are rich, and are easy to be polluted by putrefying microorganisms to generate putrefying substances in the processing process, and the putrefying bacteria mainly need aerobic bacteria, so the method selects facultative anaerobes and adopts an oxygen-isolated biological fermentation mode to deodorize. In order to ensure that the target bacteria can be rapidly propagated and deodorized, according to the characteristics of high content of nitrogen sources such as protein, amino acid and the like and low content of carbon sources such as saccharides and the like in the shellfish meat, anaerobic microbial fermentation is carried out according to the following method: adding fermentation liquor into the shellfish meat after the ozone sterilization treatment, wherein the mass ratio of the shellfish meat subjected to the ozone sterilization and deodorization to the fermentation liquor is 1: 0.5-1, preferably immersing the shellfish meat, placing the shellfish meat in a fermentation tank or a fermentation bag or other biological fermenters capable of being filled with gas and sealed, inoculating the lactobacillus paracasei subspecies paracasei RP38 lactobacillus and saccharomyces cerevisiae JJ4 in a ratio of 3-5: 1 by using composite bacteria, wherein the inoculation amount is 1-5% (v/w), and filling CO into the inoculated fermentor₂Or N₂And (3) isolating oxygen and sealing, and performing anaerobic microbial fermentation at 25-30 ℃ for 3-6 hours to obtain the shellfish meat raw material which has no obvious fishy smell and certain flower and fruit fermentation fragrance and is used for subsequent deep processing.

The preparation method of the fermentation liquor comprises the following steps: preparing sugar water with the mass concentration of 1-2% by using white granulated sugar, adjusting the pH value to 4.5-4.6 by using citric acid, sterilizing and cooling for later use.

The technical scheme of the invention has the following beneficial effects:

1. the microbial fermentation of the existing aquatic products and the like generally needs to be carried out at a high temperature suitable for target microorganisms, the fermentation time needs 3-10h, and meanwhile, the aquatic products such as shellfish meat and the like have the risk of deterioration caused by the pollution of spoilage bacteria in the biological fermentation process due to the nutritional characteristics of high protein and the like. The invention adopts the advanced ozone sterilization treatment on the shellfish meat, thereby not only reducing the base number of the pollution microorganisms in the shellfish meat raw material, but also achieving the double effects of preliminary oxidation and deodorization, and simultaneously ensuring the smooth performance of the shellfish meat biological fermentation process by adopting the oxygen isolation fermentation technology in the subsequent fermentation.

2. The method mainly comprises the steps of fermenting yeast and lactic acid bacteria such as staphylococcus or lactobacillus fermentum or lactobacillus bulgaricus or streptococcus thermophilus, and carrying out composite fishy smell removal by combining activated carbon adsorption or embedding and compounding with cyclodextrin and the like, wherein the autonomously bred lactobacillus paracasei subspecies RP38 lactic acid bacteria with stronger aquatic organism fishy smell removal capability and saccharomyces cerevisiae JJ4 with fragrance production and fishy smell removal capabilities are adopted, and simultaneously, the combination of ozone oxidation and bacterium reduction and anaerobic microorganism fermentation and fishy smell removal is adopted, so that a good fishy smell removal effect can be achieved.

Detailed Description

In order to explain the technical content, the achieved objects and effects of the technical solution in detail, the following detailed description is given with reference to specific embodiments.