CN1059464C

CN1059464C - Monascus with new strain and its application

Info

Publication number: CN1059464C
Application number: CN97100246A
Authority: CN
Inventors: 丁前胜
Original assignee: Individual
Current assignee: Individual
Priority date: 1997-01-13
Filing date: 1997-01-13
Publication date: 2000-12-13
Anticipated expiration: 2017-01-13
Also published as: CN1160764A

Abstract

The present invention relates to a new bacterial strain and application thereof in the biological technical field. N8 Monascusaurntiacus (AS3.4693) was bred from curved powder of monascus ruber of a Huawei health product factory in the jiangsu province which belongs to a Yunxi farm of a military area in the jiangsu province, and was preserved in the common micro-organism center of a micro organism strain management committee in China in August 8th, 1996, and has the registration number which is CGMCC No. 0285. In the present invention, high ethyl caproate is produced by using the new bacterial strain which is cultivated, fermented, distilled, filtered, etc. The new bacterial strain of the present invention has the advantages of stable biochemical, physical and chemical characteristics, easy preservation, low requirement for a culture medium, strong esterification capacity, large selectivity of fermentation conditions, simple technology and no pollution, solves the problems that qu wine has the disadvantages of high cost and low quality, and is favorable for human health.

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 ethyl hexanoate 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 ethyl hexanoate flavouring wine.

Technical scheme of the present invention is: monascus with new strain N ₈(Monascus aurntiacus):

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.

When 1-2 grew on the G25N substratum, bacterium colony spread expansion, and aerial hyphae is very abundant, was closely knit carpet-like, and apricot yellow to apricot pink look, back side cologne earth look has haematochrome to secrete in substratum.

1-3 is on the potato substratum, and aerial hyphae is beautiful pink.

1-4 microscopy form is a mycelium every, branch, and 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 this bacterial strain biochemistry, physicochemical property are stable, and adaptability is good, and refrigerator can be stored, and is less demanding to substratum; Its esterification 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.

Fig. 1 is a process flow sheet of the present invention.

Embodiment 1:

Monascus with new strain bacterium N ₈(Monascus aurntiacus) is 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 ₈Bacterial strain.

Former strain preparation purifying: get the koji powder one little spoon that has monascus ruber, add and to fill in 10 milliliters of maltose liquid (10Be) test tubes that contain lactic acid 0.7%, alcohol 10%, 30 ℃ leave standstill cultivation 288-302 hour, will float on the liquid level cenobium down in aseptic condition and choose in the triangular flask that fills 30 ml sterile waters, are placed with the sterilization glass strain of working hard in the bottle, shook 5 minutes, smash cenobium, filter, filtrate is diluted to proper concn with the sterilization bucket that is covered with the skim cotton, 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:

To (use the 500ml triangular flask for examination bacterium inclined-plane kind switching triangular flask, every bottled 50 gram wheat brans, after sterilizing 40 minutes under 1.5 pounds of conditions of high-pressure sterilizing pot, be cooled to inoculation below 40 ℃), after cultivating 48 hours under 40 ℃ ± 1 the condition, transfer to again after cultivating 96 to 120 hours under 40 ℃ ± 2 conditions, do the esterification fermentation routinely and detect.

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, tested in different substratum He under the different fermentations phase condition repeatedly through nearly 3 years, strong, the good stability of selected at last adaptability produces high bacterial classification 1 strain of ester rate, is delivered to the Chinese Academy of Sciences through identifying and fixed life is N ₈(Monascus aurntiacus) monascus bacterium.

Produce the method for high caproic acid flavouring wine:

Slant medium (%):

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

(2) G25N (%): NaNO0.3, K ₂PHO ₄0.1, KCL0.05, MgSO ₄.7H ₂O0.05, FeSO ₄.7h ₂O0.01, yeast extract 0.5, agar-agar 1.5, glycerine 23, fixed molten 100 milliliters, pH value 6.5-6.7;

(3) (%): yeast extract paste 0.5, glucose 1, agar-agar 2, sucrose 1, fixed molten 100 milliliters of wheat bran leach liquor, PH is worth naturally.

Secondary seed medium: high-quality wheat bran 100 grams, 600 milliliters in tap water, glucose 10 grams, after above-mentioned substance fully mixed thoroughly, branch will be 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 incubator to cultivate then 96-120 hour, treats can do after mycelia is covered with the secondary kind and uses.

Solid koji is cultivated: get and be sub-packed in koji tray after the 10kg wheat bran meets second class inoculum 3-5%, thickness is 2cm, the layering ventilating fermentation, leavening temperature is 35-38 ℃, cultivate after 96 hours, wheat bran Qu Changman mycelia also incarnadine occurs, and fermentation can stop, and dry (temperature should below 40 ℃) pulverize, and is crude zyme preparation.

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

Crude zyme preparation after pulverizing added in 5% ratio transfer in the mixed solution of 20-25 degree tail wine and yellow water, the acidity of 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.

Claims

1. monascus with new strain N ₈(Monascus aurntiacus) now is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center, and its numbering of registering on the books is: CGMCCNo.0285.

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 to produce with bent;

2-1 produces with bent the pulverizing and is crude zyme preparation, crude zyme preparation is added in 5% ratio transfer in the mixed solution of 20-25 degree tail wine and yellow water, and the acidity of 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;

Fermented liquid is distilled routinely, collect the above wine liquid of 60 degree and be high ethyl hexanoate flavouring wine, ester content can reach more than the 400mg/100ml, and the following surplus wine of 60 degree can be made lower whorl fermentation esterification mother liquor.

3. 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 to produce with bent;

3-1 the present invention also can carry out the solid fermentation high-ester flavouring wine, be about to poorly at the bottom of the pond to add 3% crude zyme preparation of throwing flow vector and to produce koji powder and drop into simultaneously and mix thoroughly into the pond, treat fermentation ends, play poor distillation separately, can contain the above high-ester flavouring wine of ethyl hexanoate 300mg/100ml, by the 5% adding crude zyme preparation of throwing flow vector, can improve ethyl hexanoate content in addition.

4. according to the production method of claim 2 or 3 described high ethyl hexanoate flavouring wine, it is characterized in that slant medium comprises:

4-1.MEA (%): malt extract 2, peptone 0.1, glucose 2, agar 1.5, pH value 5.1-5.4;

4-2.G25N (%): N _aNO ₃0.3, K ₂HPO ₄0.1, KCL0.05, MgSO ₄.7H ₂O0.05, F _eSO ₄7H ₂O0.01, yeast extract 0.5, agar 1.5, glycerine 23, pH value 6.5-6.7;

4-3. (%): yeast extract paste 0.5, glucose 1, agar 2, sucrose 1, fixed molten 100 milliliters of wheat bran leach liquor, PH is worth naturally.