CN1160764A

CN1160764A - Monascus with new strain and its application

Info

Publication number: CN1160764A
Application number: CN 97100246
Authority: CN
Inventors: 丁前胜
Original assignee: Individual
Current assignee: Individual
Priority date: 1997-01-13
Filing date: 1997-01-13
Publication date: 1997-10-01
Anticipated expiration: 2017-01-13
Also published as: CN1059464C

Abstract

The present invention relates to biological technology field. The present invention breeds from Monascus powder new strain N8Monascusaurntiacus (AS3.4693) which is preserved in Common Microbe Center of Chinese Microbe Strain Management Committee in Aug. 8.1996 in the number of CGMCC No.0285. By using the new strain, the present invention produces ethyl perhexanoate through culture, fermentation, distillation, filtering and other processes. The new strain has stable physical and chemical properties to preservation, no excessive demands to culture medium and fermentation condition, strong esterifying capacity and no pollution. The present invention also solves the problem of high cost and low quality of wine yeast.

Description

Monascus with new strain and application thereof

The present invention relates to a kind of new bacterial strain and application thereof, belong to microbial technology field.

Ethyl hexanoate is the main body aroma-producing substance of aroma daqu liquor.Before the present invention makes, main improve ethyl hexanoate content in the bent wine by the prolongation yeast phase.This method gained caproic acid second fat content is very little, and influences the yield of liquor, bent wine usually occurs summer and falls to arrange phenomenon.

Purpose of the present invention will overcome above-mentioned defective exactly, extracts and obtains monascus with new strain and produce high caproic acid second fat.

Technical scheme of the present invention is: when monascus with new strain N8Monascusaurntiacus (AS3.4693): 1-1. grew on malt extract medium and MEN substratum, the bacterium colony yellow spread expansion, and a small amount of fold is arranged; The abundant velvet shape that is of aerial hyphae; It is light yellow that mycelium is initially, and is orange, the shallow chocolate in back after aging.1-2. when growing on the G25N substratum, bacterium colony spreads expansion, aerial hyphae is very abundant, is closely knit carpet-like, and apricot yellow to apricot pink look, back side cologne earth look has haematochrome to secrete in substratum.1-3. on the potato substratum, the beautiful pink of aerial hyphae.1-4. the microscopy form is that mycelium is separated, branch, wall is more smooth, and diameter 3-7 micron includes red oil droplet; The quilt device is orange red, spherical, diameter 40-75 micron, and thecaspore is smooth colourless, and oval 5 * 7 * 4.5-5 micron is avette 4.5 * 7 microns.2. the production method of high ethyl hexanoate flavouring wine, it is characterized in that adopting new bacterial strain, after the slant culture inoculation, under temperature 30-32 ℃ condition, cultivate after 72-96 hour, the switching eggplant-shape bottle is cultivated, and after cultivating 72-96 hour under the 30-32 ℃ of condition, switching triangular flask bran mass is cultivated, after cultivating 96-120 hour under the 32-36 ℃ of condition, the bent substratum of switching ventilating distiller's yeast or curtain was cultivated 72 hours under 35-38 ℃ of condition, was and produced with bent.

Advantage of the present invention and effect are that bacterial strain biochemistry, physicochemical property are stable, and adaptability is good, and refrigerator can be stored, and is less demanding to substratum; Its fat power is strong, and the fermentation condition selectivity is big, and technology is simple and pollution-free; Solved and fallen to arrange phenomenon summer, also solved bent wine cost height, a ropy difficult problem, and helped HUMAN HEALTH, saved food.

Embodiment 1:

Monascus with new strain bacterium N8Monascus aurntiacus (AS3.4693) is a fungi, is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center on August 8th, 1996, and that registers on the books is numbered: CGMCC No.0285.

The bacterial classification source: bacterium source of the present invention has the bent powder of monascus ruber in Yun Xi farm, the military region, Jiangsu Province China prestige healthcare products factory.Get 6 parts of bent powder, after difference purifying enlarged culturing, make application test in testing laboratory, find to have product ester (ethyl hexanoate) bacterial strain that ability is the highest, purified again screening, production test obtain N at last ₈(AS3.4693) bacterial strain.

Former strain preparation and purifying: get the koji powder one little spoon that has monascus ruber, add in the test tube fill 10 milliliters of maltose liquid (10Be) that contain lactic acid 0.7%, alcohol 10%, 30 ℃ leave standstill cultivation 288-302 hour, to float on the liquid level cenobium down in aseptic condition chooses in the triangular flask that fills 30 ml sterile waters, be placed with some sterilization granulated glass spherees in the bottle, shook 5 minutes, smash cenobium, filter with the sterilization bucket that is covered with the skim cotton, filtrate is diluted to proper concn, be coated with flat board, cultivated 5 days in 35 ℃.The bacterium colony that bigger, medium, the less and color of choosing colony is consistent moves into the maltose inclined-plane respectively and uses for screening.

Primary dcreening operation:, make the fermentation simultaneous test, the primary dcreening operation step with the production Daqu with above isolated strains 36 strains:

The result shows, produces 6 above strains of ethyl hexanoate 300mg/100ml, wherein produces 12 strains that ethyl hexanoate is lower than 100mg/100ml, and all the other are all about 200mg/100ml.

Multiple sieve: esterification power is further purified at the bacterial strain more than 300, and the simultaneous test of still fermenting, esterification power is in 3 strains more than 400.

Original seed is selected: after 3 strain bacterial classifications after will sieving are again cultivated respectively, produce the curtain song.Enter and produce the contrast fermentation test, in different substratum He under the different fermentations phase condition, tested repeatedly through nearly 3 years, strong, the good stability of selected at last adaptability, produce high bacterial classification 1 strain of ester rate, be delivered to the Chinese Academy of Sciences and be N8Monascus aurntiacus monascus bacterium AS3.4693 through identifying and deciding life.

Produce the method for high ethyl hexanoate flavouring wine:

Minimum medium (%):

(1) malt extract 2, peptone 0.1, glucose 2, agar 1.5, fixed molten 100 milliliters, pH value 5.1-5.4.

(2) NaNO ₃0.3, K ₂HPO ₄0.1, KCL0.05, MgSO ₄.7H ₂O0.05, FeSO ₄.7H ₂O0.01, yeast extract 0.5, agar 1.5, glycerine 23, fixed molten 100 milliliters, pH value 6.5-6.7.

Secondary seed medium: high-quality wheat bran 100 grams, 600 milliliters in tap water, glucose 10 grams, after above-mentioned substance fully mixed thoroughly, be sub-packed in 500 milliliters of triangular flasks of band tampon every bottle 50 gram, after 1.5 pounds of high-pressure sterilizing pots were sterilized 40 minutes, taking-up is cooled to below 40 ℃ and inoculates under aseptic condition, places 32-36 ℃ of substratum to cultivate then 96-120 hour, treats can do after mycelia is covered with the secondary kind and uses.

Solid koji is cultivated: get the 10kg wheat bran and be sub-packed in koji tray, thickness is 2cm, the layering ventilating fermentation, leavening temperature is 35-38 ℃, cultivates after 96 hours, and wheat bran Qu Changman mycelia also incarnadine occurs, fermentation can stop, and dry (temperature should below 40 ℃) pulverize, and is crude zyme preparation.

The slant tube bacterium of monascus bacterium was transferred inclined-plane 30-32 ℃ of following cultivation of eggplant type bottle after 72-96 hour under the aseptic condition, apricot pink look appears in substratum and lawn, the triangular flask bran mass of transferring is again cultivated, after cultivating 96-120 hour under the 32-36 ℃ of condition, change the bent cultivation of aerated koji making or curtain, culture temperature is 35 ℃ ± 3, and ventilation is looked bent Wen Erding, cultivate after 72 hours, wheat bran incarnadine occurs and gets final product cryodrying, pulverizing.

Crude zyme preparation after hand over pulverizing adds in 5% ratio and transfers in the mixed solution of 20-25 degree tail wine and yellow water, and the acidity in the mixed solution is 1%, stirs evenly to seal to be placed on that esterification was a fermentation ends in 7 days under 35 ℃ ± 3 conditions.

With the fermented liquid distillation, to collect the above wine liquid of 60 degree and be high ethyl hexanoate flavouring wine, ester content reaches more than the 400mg/100ml.The following surplus wine of 60 degree can be made lower whorl fermentation esterification mother liquor (substratum).

Embodiment 2:

In order to simplify technology, can utilize fermentation vat to carry out the solid fermentation high-ester flavouring wine, concrete grammar is: poor 3% crude zyme preparation that adds the throwing flow vector at the bottom of the pond is dropped into simultaneously with the production koji powder mix thoroughly into the pond, treat fermentation ends (being generally 45-60 days), play poor distillation separately, can contain the above high-ester flavouring wine of ester 300mg/100ml.5% adding crude zyme preparation by throwing flow vector can improve ethyl hexanoate content, reduces the content of ethyl lactate, thereby improves the output and the quality of wine.

Technical process (as figure).

Claims

1. monascus with new strain N8Monascus aurntiacus (AS3.4693), it is characterized in that: when 1-1. grew on malt extract medium and MEN substratum, the bacterium colony yellow spread expansion, and a small amount of fold is arranged; The abundant velvet shape that is of aerial hyphae; It is light yellow that mycelium is initially, and is orange, the shallow chocolate in back after aging.1-2. when growing on the G25N substratum, bacterium colony spreads expansion, aerial hyphae is very abundant, is closely knit carpet-like, and apricot yellow to apricot pink look, back side cologne earth look has haematochrome to secrete in substratum.1-3. on the potato substratum, the beautiful pink of aerial hyphae.1-4. the microscopy form is that mycelium is separated, branch, wall is more smooth, and diameter 3-7 micron includes red oil droplet; The quilt device is orange red, spherical, diameter 40-75 micron, and thecaspore is smooth colourless, and oval 5 * 7 * 4.5-5 micron is avette 4.5 * 7 microns.

2. the production method of high ethyl hexanoate flavouring wine, it is characterized in that adopting the described new bacterial strain of claim 1, after the slant culture inoculation, under temperature 30-32 ℃ condition, cultivate after 72-96 hour, the switching eggplant-shape bottle is cultivated, and after cultivating 72-96 hour under the 30-32 ℃ of condition, switching triangular flask bran mass is cultivated, after cultivating 96-120 hour under the 32-36 ℃ of condition, the bent substratum of switching ventilating distiller's yeast or curtain was cultivated 72 hours under 35-38 ℃ of condition, was and produced with bent.

3. the production method of high ethyl hexanoate flavouring wine according to claim 2 is characterized in that slant medium comprises: 3-1. MEA (%): malt extract 2, peptone 0.1, glucose 2, agar 1.5, pH value 5.1-5.4.3-2. G25N (%): NaNO ₃0.3, K ₂HPO ₄0.1, KCL0.05, MgSO ₄.7H ₂O0.05, FeSO ₄7H ₂O0.01, yeast extract 0.5, agar 1.5, glycerine 23, pH value 6.5-6.7.3-3. yeast extract paste 0.5, glucose 1, agar 2, sucrose 1, wheat bran leaches certainly molten 100 milliliters, and PH is worth naturally.