CN115521848A - Method for producing rose vinegar based on mixed starter propagation of monascus black koji rhizopus - Google Patents

Abstract

The invention discloses a method for producing rose vinegar by mixed starter propagation based on monascus black koji rhizopus, which comprises the steps of rice soaking and washing, rice steaming, starter propagation inoculation, fermentation, after-ripening and squeezing; the inoculation starter propagation step comprises the following steps: selecting pure strains of red yeast, black yeast, rice yeast and rhizopus as fermentation strains, and inoculating the fermentation strains into the cooled corresponding cooked rice in a proportion of 1 per mill; the fermentation step comprises: during feeding, mixing the red yeast finished koji, the black koji finished koji, the rice koji finished koji and the rhizopus finished koji according to the ratio of 1. The method not only finds out the types of the mould from practice, but also calculates the adding proportion of each pure variety by using a dynamic balance model of saccharification-alcoholic fermentation-acetic acid fermentation, omits the traditional step of flowering, solves the technical problem of quality difference among finished products in cylinders in the traditional rose vinegar production, shortens the production period, is beneficial to large-scale industrial production, and improves the production efficiency of the rose vinegar.

Description

Method for producing rose vinegar based on mixed starter propagation of monascus black koji rhizopus

Technical Field

The invention relates to a method for producing rose vinegar, in particular to a method for producing rose vinegar based on mixed starter propagation of red yeast black koji rhizopus oryzae.

Background

The traditional Zhejiang rose vinegar production method comprises the following basic steps: soaking and washing rice, steaming, cooling, making nest, growing flower, adding water, fermenting, ageing, squeezing, sterilizing and blending. The method for promoting the growth of the rice comprises a mixed saccharification process of allowing wild strains (including aspergillus oryzae, aspergillus niger, monascus and the like which grow in the process of promoting the growth of the rice, and yeasts, lactic acid bacteria, acetic acid bacteria and the like which fall in the process of turning over the rice jar) to fall on the surface of the rice in the rice jar. Therefore, the rose-scented vinegar belongs to multi-bacterium co-fermentation.

The rose vinegar brewed by natural mixed strains (including mould, yeast and acetic acid bacteria) has the characteristics of light rose color, soft and refreshing sourness and slightly fresh and sweet taste. However, the degree of colony growth and saccharification of rice varies greatly from jar to jar due to fermentation with wild fungi:

firstly, the rose vinegar is brewed by adopting natural mixed strains, the proper reproduction proportion of various moulds in the rice cannot be artificially controlled, and a considerable part of the rice cannot be saccharified well, so that the quality difference among finished products in each jar is caused; secondly, the rose vinegar is brewed by adopting natural mixed strains, natural fermentation which lasts for half a year (5-11 months of brewed vinegar) is needed, manual operation is mainly used in nest building and cylinder turning operation, and the defects of long production period, unstable quality, low yield, strong seasonality and the like exist in the flowering process. The lower industrial production level of the method can not meet the market demand, and the food safety has hidden troubles.

In order to improve the edible safety of the rose vinegar, shorten the fermentation period, stabilize the quality of the rose vinegar and realize mechanized production, the inventor has previously applied a pure fermentation technology to the production of the rose vinegar, but still has more problems which are not solved:

firstly, a reasonable pure variety and a reasonable proportion thereof cannot be found, so that the problem of unbalanced trilateral (saccharification, alcoholization and acetification) fermentation proportion exists in the production process of the rose vinegar, and the high-alcohol low-acid vinegar exists, namely the alcoholic strength of the fermentation liquor is kept between 8 and 9vol percent, and the acidity is kept about 3g/mL for a long time. Through research of the inventor for years, the method has the following discovery that: this phenomenon is mainly due to the fact that the diastase activity in the fermentation broth is too high, and the alcoholization process is too fast (E, F equation has higher reaction speed); the acetic acid process is too slow (the reaction speed of the G equation is low), the degree of the acetic acid process cannot be kept up to that of the reaction process of the G equation, and the growth of acetic acid bacteria is inhibited by the quickly accumulated alcoholic strength;

secondly, the inventor also finds that: the unreasonable pure fermentation proportion can also cause the phenomenon of 'jar falling', namely, the activity of saccharifying enzyme in fermentation liquor is low, starch gelatinization is not thorough (the equation E is slowly carried out), so that vinegar mash in rose vinegar fermentation is not layered (high-molecular dextrin content causes high viscosity of mash), sugar is less formed, alcohol fermentation is too slow (the equation F is slow), wild yeast is quickly propagated to cover the liquid surface, so that acetic acid bacteria cannot obtain enough oxygen, the acetic acid bacteria are gradually killed (reaction of the equation G is inhibited), and the phenomenon of turbidity and mucus of the fermentation mash is further caused. How to determine the proper addition ratio of each pure species is a technical problem which besets the technical personnel in the field.

(C ₆ H ₁₀ O ₅ )nH ₂ O→xC ₆ H ₁₂ O ₆ +yH ₂ O………………E

C ₆ H ₁₂ O ₆ →2C ₂ H ₅ OH+2CO ₂ ………………F

C ₂ H ₅ OH+O ₂ →CH ₃ COOH+H ₂ O………………G

Disclosure of Invention

The invention aims to provide a method for producing rose vinegar by mixed koji making based on red yeast black koji rhizopus oryzae, which not only finds out the variety of the mold from practice, but also calculates the adding proportion of each pure variety by a dynamic balance model of saccharification-alcoholic fermentation-acetic acid fermentation. The method omits the traditional step of flowering, solves the technical problem of quality difference between finished products in cylinders in the traditional rose vinegar production, shortens the production period, is beneficial to large-scale industrial production, and improves the production efficiency of the rose vinegar.

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

a method for producing rose vinegar based on mixed starter propagation of red yeast black starter rhizopus oryzae comprises the steps of rice soaking and washing, rice steaming, starter propagation inoculation, fermentation, after-ripening and squeezing; the inoculation starter propagation step comprises the following steps:

step a, selecting pure strains of red yeast rice (As 3.972), black yeast (Hu Niang 3.041), rice koji (As 3.042) and rhizopus (Hu Niang 3.866) As fermentation strains, inoculating the fermentation strains into corresponding cooled cooked rice (also called rice) according to the proportion of 1 per mill, controlling the moisture content of the cooked rice to be 70%, controlling the relative air humidity of a koji room to be 94-96%, culturing the black yeast at 30 ℃ for 60h, culturing the red yeast rice, the rice koji and the rhizopus at 32 ℃ for 72h, and controlling the moisture content of the finished yeast to be 65-70%;

the fermentation step comprises:

and step b, mixing the red yeast, the black yeast, the rice yeast and the rhizopus yeast according to the proportion of 1.

The rice is late long-shaped rice, the rice is soaked at room temperature for 12 days at 15 ℃, soaked for 10 days at 20-25 ℃, soaked for 7 days at the temperature of more than or equal to 25 ℃, the pH value of rice soaking water is controlled to be 3.0-3.2 when the rice soaking is finished, the rice (cooked rice) yield is controlled to be 200-220% when the rice is steamed, the cooked rice is inoculated after being cooled to 39-41 ℃ on a bamboo plaque or a stainless steel sieve plate, and the thickness of the cooked rice layer is controlled to be 9.5-10.5 cm.

C, after flushing the tank and discharging water, controlling the fermentation temperature to be 30-33 ℃, manually or mechanically turning the tank once every other day, wherein the medium-frequency high-stirring intensity in the early period, the medium-frequency high-stirring intensity in the middle period and the low-frequency low-stirring intensity in the later period are used for turning the tank, and after 77-80 days, adding 1.5-2.5% of salt to finish the fermentation.

In the step b, the yeast materials are mixed in a temperature-controlled fermentation tank or jar; and c, adding clean clear water according to the proportion that the total koji is water =1:3, uniformly mixing, and then covering a grass cylinder cover for carrying out rose vinegar fermentation.

The mixing proportion of the red yeast finished koji, the black koji finished koji, the rice koji finished koji and the rhizopus finished koji =1 in the step b is obtained according to the following steps:

the proportion relation of various enzyme activities in rice grains at the late flowering stage of traditional rose vinegar is as follows:

alpha-amylase beta-amylase protease equal to 1,

the enzyme activity value (U/g) of each mould is referred to as:

the proportions of red yeast rice, black yeast, rice koji and rhizopus are A, B, C, D respectively, and the proportions are as follows:

(1)80A+400B+200C+200D＝400A+600B+600C+1500D

(2)2*(80A+400B+200C+200D)＝350A+1100B+1000C+200D

decomposing to obtain:

(3)24A+20C+110D＝20B

(4)11A+80C＝10B

substituting values of C =1,D =1 by an exhaustion method to obtain the addition ratio of the strains, which is as follows, that the addition ratio of the red yeast finished koji, the black koji finished koji, the rice koji finished koji, the rhizopus finished koji = 1.

Compared with the prior art, the invention has the beneficial effects that:

on the basis of proper strain selection and addition proportion, the koji-making conditions (temperature, humidity and koji-making time) can generate great influence on various enzyme activities of the koji-making. The invention considers the influence of temperature and koji-making time on the activity of koji-making enzyme. Aspergillus niger, a major industrial strain for the production of saccharifying enzymes, grows faster than other strains and has a high ability to saccharify and decompose proteins. When the aspergillus niger is used for producing amylase, the temperature is usually selected to be 30 ℃ for culturing for 60h, and the saccharifying power reaches 633U/g; the production of the protease is usually carried out by culturing at 30 ℃ for 48h, and the activity of the protease can reach 1200U/g. Through multiple experiments and enzyme activity determination, the culture time and temperature for preparing the black koji are optimally selected to be 60h and 30 ℃. In the starter propagation production of soy sauce by aspergillus oryzae, bean pulp is used as a raw material for starter propagation, and the activity of neutral protease can reach 2000-3000U/g. Culturing at 32 deg.C for 72 hr on rice, and the saccharifying enzyme activity can reach 860U/g and protease activity can reach 1000U/g. The growth temperature and humidity conditions of the red yeast are similar to those of the rice koji, the red yeast has higher liquefaction and saccharification force and certain protease activity, and after the red yeast is cultured for 72 hours at 32 ℃, the activity of the saccharification enzyme reaches 800U/g, and the activity of the protease reaches 350U/g. The rhizopus has strong saccharifying power, and the activity of the saccharifying enzyme can reach 1520U/g after culturing at 32 ℃ for 70 hours. The invention adopts microbial strains which have higher activity of the needed starch liquefying enzyme and saccharifying enzyme and moderate activity of protease.

Secondly, researches show that the dominant bacteria in the natural flowering process comprise mold and yeast, mainly comprising rhizopus, aspergillus and penicillium, saccharomyces boulardii, rhodosporidium toruloides, rhodotorula glutinis, kluyveromyces marxianus and the like. The mould has main advantages in the early stage of the flowering process, the growth is vigorous, the temperature in the fermentation tank is close to 40 ℃, and the fermentation tank is suitable for the growth of the mould and bacteria. In previous studies, pure fermentation technology has been applied to the production of rose vinegar, but many problems remain unsolved. Firstly, the unreasonable pure seed ratio has the problems of unbalanced trilateral (saccharification, alcoholization and acetification) fermentation ratio and the like, so that the high-alcohol and low-acid vinegar exists, namely the alcoholic strength of the fermentation liquor is kept between 8 and 9vol percent, and the acidity is kept about 3g/mL for a long time. The phenomenon is mainly caused by that the saccharifying enzyme in the fermentation liquor has overhigh activity, the alcoholization is carried out too fast, the acetification process is carried out too slow, the alcoholization degree cannot be kept up to the former degree, and the generated alcoholicity inhibits the growth of acetic acid bacteria. Secondly, the unreasonable proportion of pure fermentation may cause the phenomenon of 'jar falling', i.e. the saccharifying enzyme activity in the fermentation liquor is low, starch gelatinization is not thorough, so that vinegar mash is not layered in rose vinegar fermentation, alcohol fermentation is too slow, wild yeast is propagated in the fermentation liquor in large quantities to cover the liquid surface, acetic acid bacteria cannot obtain enough oxygen and die in large quantities, and the fermentation mash has the phenomena of turbidity and mucus (commonly called 'tung oil vinegar'). The invention adopts a mode of independent starter propagation mixed fermentation, has proper strain addition proportion, follows the proportion of each enzyme activity in the traditional fermented rose vinegar, ensures that the added strains are in an advantageous state in the whole fermentation process of the rose vinegar, does not generate the phenomena of 'high-wine low-acid vinegar' and 'tung oil vinegar', and improves the safety of the production of the rose vinegar.

And finally, the leavening is replaced by the starter propagation, so that the production efficiency is greatly improved. In the traditional fermentation mode, the flowering is carried out in a jar, in order to easily control the flowering and the fermentation temperature, the adding amount of rice in each jar is controlled at 100kg, the scale is small, the stacking thickness exists, surface hypha is difficult to penetrate into the rice, the gelatinization of a substrate is uneven and incomplete, and the whole enzyme activity is limited. The traditional rose vinegar production adopts a pure manual fermentation mode, and the labor intensity is high. The automatic starter propagation can effectively reduce the labor intensity in the production process and improve the production efficiency. By adopting starter propagation, hypha can grow on each grain of rice, the mould can thoroughly decompose the substrate, the production efficiency of the rose vinegar is improved, the yield of the obtained rose vinegar is 6.10. The new process of replacing the flower formation with the pure-breed starter propagation greatly improves the mechanization degree of the production and reduces the energy consumption.

Drawings

FIG. 1 is a schematic view of the production process of the present invention.

Detailed Description

In order to make the technical solution of the present invention clearer, the present invention is described in detail below with reference to fig. 1. It should be understood that the detailed description and specific examples, while indicating the preferred embodiment of the invention, are intended for purposes of illustration only and are not intended to limit the scope of the invention.

The invention relates to a method for producing rose vinegar by mixed starter propagation based on monascus black starter rhizopus, which comprises the steps of rice soaking and washing, rice steaming, inoculation starter propagation, fermentation, after-ripening and squeezing;

preferably, the rice is late long-shaped rice, the rice is soaked at room temperature for 12 days at 15 ℃, the rice is soaked for 10 days at 20-25 ℃, the rice is soaked for 7 days at the temperature of not less than 25 ℃, the pH value of rice soaking water is controlled to be 3.0-3.2 when the rice soaking is finished, the rice (cooked rice) rate is controlled to be 200-220% when the rice is steamed, the cooked rice is inoculated after being cooled to 39-41 ℃ on a bamboo plaque or a stainless steel sieve plate, and the thickness of the cooked rice layer is controlled to be 9.5-10.5 cm;

the inoculation starter propagation step comprises the following steps:

step a, selecting pure strains of red yeast As 3.972, black yeast Hu brewing 3.041, rice koji As 3.042 and rhizopus Hu brewing 3.866 As fermentation strains, inoculating the fermentation strains into corresponding cooled cooked rice (also called rice) according to the proportion of 1 per mill, controlling the water content of the cooked rice to be about 90 percent, controlling the relative air humidity of a yeast room to be 94-96 percent, culturing the black yeast at 30 ℃ for 60h, culturing the red yeast, the rice koji and the rhizopus at 32 ℃ for 72h, and controlling the water content of the finished yeast to be 65-70 percent;

the fermentation step comprises:

step b, mixing the red yeast finished koji, the black koji finished koji, the rice koji finished koji and the rhizopus finished koji according to a ratio of 1;

c, adding the mixed yeast material into a temperature-controlled fermentation tank or jar, adding clean water according to the proportion of the total yeast, namely water =1:3, uniformly mixing, and then covering a grass jar cover for rose vinegar fermentation;

the fermentation (alcoholization, acetification), after-ripening and squeezing steps can all adopt a conventional method, preferably, after flushing a jar and discharging water, the fermentation temperature is controlled to be 30-33 ℃, the early-stage stirring frequency is 3 times/2 weeks when turning over the jar, the stirring speed is 115-135 r/min, the stirring time is 3-5 min, a grass cover is opened during stirring, the middle-stage stirring frequency is 2-4 times/week, the stirring speed is 105-115 rpm, the stirring time is 1-3 min, the grass cover is opened during stirring, the later-stage stirring frequency is 1 time/week, the stirring speed is 90-100 rpm, the stirring time is 1-3 min, the grass cover is covered during stirring (specifically, the pneumatic/fermented mash backflow/mechanical stirring is carried out in a mode of CN109971605A, CN108559691A, CN107916209A, the temperature, the oxygen supply and the feed liquid are fully mixed during the fermentation process is controlled), after 77-80 days, 1.5-2.5% of salt is added, the fermentation temperature is reduced to be below 25 ℃, and the action of acetic acid bacteria is stopped to finish the fermentation; then storing for 1 month at normal temperature, squeezing, and sterilizing to obtain the final product.

The mixing ratio of the red yeast finished koji to the black koji finished koji to the rice koji finished koji to the rhizopus finished koji =1 is as follows:

the proportion relation of the activity of each enzyme in the later stage of the traditional rose vinegar flowering can be expressed as follows:

the proportion relation of alpha-amylase, beta-amylase and protease is equal to 1.

The enzyme activity value (U/g) of each mould is referred to as:

the proportions of red yeast rice, black yeast, rice yeast and rhizopus are A, B, C, D respectively, which comprises:

(1)80A+400B+200C+200D＝400A+600B+600C+1500D

(2)2*(80A+400B+200C+200D)＝350A+1100B+1000C+200D

decomposing to obtain:

(3)24A+20C+110D＝20B

(4)11A+80C＝10B

substituting the numerical values of C =1 and D =1 by an exhaustion method to obtain the following addition ratios of the strains, namely, the ratio of monascus finished koji to black koji to rice koji to rhizopus finished koji = 1.

The comparison experiment of the new pure fermentation process and the traditional rose vinegar production process comprises the following steps: 1 test conditions and environments

The site: wuxing, huzhou, laoheng and brewing, inc.;

time: 2021.05-2021.09;

laboratory conditions: 12 fermentation test tanks (1 m) ³ One), 2 traditional fermentation pottery jars (1 m) ³ And/or) both fermentations have the same fermentation room temperature, air relative humidity, and microbial community status in contact with them during fermentation is also substantially the same.

2 test procedure

(1) No. 5/21 steaming, cooling to room temperature, and adding red yeast rice, rice koji, black koji, and Rhizopus koji. The starter propagation time is 60 to 72 hours.

Setting 2 traditional fermentation tanks as a control, feeding 50kg of rice/tank in 25 days in 5 months, building a nest, covering a grass jar cover, and carrying out flowering at natural temperature (about 26-29 ℃). The time spent was 15 days.

Mixing the yeast and the rice in 8 test jars according to the proportion, mixing the finished yeast: water (w/w) =1:3 flushing the tank and discharging water, and carrying out saccharification, alcohol fermentation and acetic acid fermentation. 2 traditional fermentation vats are according to rice: water =1:3 water flush.

The alcohol content can slowly rise to 6-7% within a week after flushing and discharging water, the acidity is slowly accumulated to about 1.0-1.2 g/100ml, and the change of substances in the jar is gradually changed into trilateral fermentation mainly based on acetic acid fermentation from starch saccharification and alcohol fermentation.

After water is discharged from the flushing tank in step (5), all fermentation tanks ferment in natural environment according to the traditional production mode, and refer to the mode of CN109971605A, CN108559691A, CN107916209A for turning over the tanks.

And (6) when the acidity reaches more than 5-6%, adding 2.5% of salt to terminate the fermentation, and ending the acetic fermentation. After-ripening is continued for 30 days at normal temperature.

The rose vinegar fermentation period of (7) based on the traditional flowering process is about 120-130 days, and the rose vinegar fermentation period based on the pure-breed starter propagation of the invention is about 100-115 days.

3 results of the test

Pilot scale production test results

Note: during the test, (1) and (3) # are other proportion test groups, and (2) # is the proportion of the patent. From the measured results, the rose vinegar sample ((2) #) produced by adopting the pure breeding technology completely meets the requirements of national standard GB/T18187-2000 for brewing vinegar; compared with Zhejiang rose vinegar group standard (Zhejiang rose rice vinegar T/ZJFA 002-2019), the sensory characteristics of the test sample such as color, aroma, taste, body form and the like also completely meet the sensory requirements of Zhejiang rose vinegar. The rose vinegar yield obtained by the test group (2) # is higher than that of a control and other mixture ratios, and the ton grain yield is improved to about 6.10 from the traditional 5.13.

Claims (5)

1. A method for producing rose vinegar based on mixed starter propagation of red yeast black koji rhizopus oryzae is characterized by comprising the steps of rice soaking and washing, rice steaming, inoculation starter propagation, fermentation, after-ripening and squeezing;

the inoculation and starter propagation step comprises the following steps:

step a, selecting pure strains of red yeast rice (As 3.972), black yeast (Hu Niang 3.041), rice koji (As 3.042) and rhizopus (Hu Niang 3.866) As fermentation strains, inoculating the fermentation strains into corresponding cooled cooked rice (also called rice) according to the proportion of 1 per mill, controlling the moisture content of the cooked rice to be 70%, controlling the relative air humidity of a koji room to be 94-96%, culturing the black yeast at 30 ℃ for 60h, culturing the red yeast rice, the rice koji and the rhizopus at 32 ℃ for 72h, and controlling the moisture content of the finished yeast to be 65-70%;

the fermentation step comprises:

and step b, mixing the red yeast, the black yeast, the rice yeast and the rhizopus yeast according to the proportion of 1.

2. The method for producing rose vinegar based on mixed koji making of red yeast black koji rhizopus oryzae according to claim 1, characterized in that: the rice is late long-shaped rice, the rice is soaked at room temperature for 12 days at 15 ℃, soaked for 10 days at 20-25 ℃, soaked for 7 days at the temperature of more than or equal to 25 ℃, the pH value of rice soaking water is controlled to be 3.0-3.2 when the rice soaking is finished, the rice (cooked rice) yield is controlled to be 200-220% when the rice is steamed, the cooked rice is inoculated after being cooled to 39-41 ℃ on a bamboo plaque or a stainless steel sieve plate, and the thickness of the cooked rice layer is controlled to be 9.5-10.5 cm.

3. The method for producing rose vinegar based on mixed koji-making of red yeast black koji rhizopus according to claim 1, which is characterized in that:

c, after flushing the tank and discharging water, controlling the fermentation temperature to be 30-33 ℃, manually or mechanically turning the tank once every other day, wherein the medium-frequency high-stirring intensity in the early period, the medium-frequency high-stirring intensity in the middle period and the low-frequency low-stirring intensity in the later period are used for turning the tank, and after 77-80 days, adding 1.5-2.5% of salt to finish the fermentation.

4. The method for producing rose vinegar based on mixed koji making of red yeast black koji rhizopus oryzae according to claim 3, characterized in that: in the step b, the yeast materials are mixed in a temperature-controlled fermentation tank or jar; and c, adding clean clear water according to the proportion of the total yeast, namely water =1:3, uniformly mixing, and then covering a grass jar cover for carrying out rose vinegar fermentation.

5. The method for producing rose vinegar based on mixed koji making of red yeast black koji rhizopus oryzae according to claim 1, wherein the mixing ratio of red yeast rice koji, black koji, rice koji and rhizopus koji =1 in step b is obtained by the following steps:

the proportion relation of various enzyme activities in the rice grains at the late flowering stage of the traditional rose vinegar is as follows:

alpha-amylase beta-amylase protease equal to 1,

the enzyme activity value (U/g) of each mould is referred to as:

the proportions of red yeast rice, black yeast, rice yeast and rhizopus are A, B, C, D respectively, which comprises:

(1)80A+400B+200C+200D＝400A+600B+600C+1500D

(2)2*(80A+400B+200C+200D)＝350A+1100B+1000C+200D

decomposing to obtain:

(3)24A+20C+110D＝20B

(4)11A+80C＝10B

substituting the numerical values of C =1 and D =1 by an exhaustion method to obtain the following addition ratios of the strains, namely, the ratio of monascus finished koji to black koji to rice koji to rhizopus finished koji = 1.

Images