CN85108564A - With black-koji mould fermentation coproduction diosgenin and citric acid - Google Patents

Abstract

With the yam that contains dioscin is raw material, with the black-koji mould fermentation, but two products of combination producing diosgenin and citric acid.By fermentation, the molecular structure of diosgenin remains unchanged, and the productive rate of diosgenin is significantly improved; By fermentation, transfer starch to citric acid and enter liquid phase, the material that drops into hydrolytic process is reduced, the consumption of acid, solvent and energy reduces, and can make the production cost of diosgenin per ton reduce about 30,000 yuan.

Description

With black-koji mould fermentation coproduction diosgenin and citric acid

The present invention is a kind of science fermentation process that is extracted diosgenin by yam.

Diosgenin is a precursor of producing multiple steroid drugs, the steroid hormone of produced worldwide, and diosgenin has drawn from more than 60%.Largest production state is a Mexico in the world, about 500 tons of annual production, and raw material variety is Dioscorea camposita and barbasco.Next is China, and raw material variety is Rhizome of Peltate Yam and Dioscorea nipponica Mak. Ningpo Yam Rhizome, and other all has a certain amount of production as states such as Guatemala, Puerto Rico, India.Estimate that it is 2400 tons that world's annual production in 1985 (not comprising China) will increase, rate of growth is 10% every year on average.

In early days, the method for producing diosgenin is a direct acid-hydrolysis method-Chinese yam sheet is put into hydrolytic decomposition pot, and with certain density sulfuric acid or hydrochloric acid catalysis hydrolysis, with organic solvent lixiviate diosgenin from hydrolyzate, sugar and starch is stayed in the waste residue and liquid then.This method hydrolysis is not thorough, and the saponin yield is low, generally reaches about 2%.

If through pre-fermentative processing, can make output improve 17～48%, yet existing pre-fermentative processing is with Chinese yam sheet water-sprinkling humidification, stacking in heaps, spontaneous fermentation, or be called from enzymic fermentation.The soaking fermentation in the pond that has, the adding exogenous enzyme that has promotes fermentation.Simple and easy to do but the science not of aforesaid method, name is said from enzymic fermentation, assorted actually bacterium fermentation, any bacterial classification in the empty G﹠W all can be participated in, and often causes and goes mouldy.Wear Chinese yam rhizomes such as dragon, shield leaf, Foochow and hollyhock leaf, mouldy back saponin must be measured height, but the fusing point reduction can obtain table-yuccagenin and yuccagenin ketone through separating, without mouldy then not having.The appearance of these two kinds of derivatives makes quality product nonconforming, and output is descended.Particularly weather condition are to big from the enzymic fermentation influence, and fermentation condition and attenuation degree are difficult to control.Certain saponin factory is because of the heavy rain of a bust makes three, 40 tons of fresh ginger slices that fermenting all mould bad, loses to extract to be worth and to be forced to outwell.Not only be difficult to promote effectively hydrolysis to improve yield from enzymic fermentation, and, make the diosgenin structural modification, change other derivatives into, reduce the product fusing point, widen molten distance, cause bigger loss often owing to go mouldy in the part.Thereby the saponin yield of most producers is still 2～2.2%.

Be the approach of seeking to improve the saponin yield He utilize starch, Rothock, J.W. etc. once used terreus MF-118 bacterial strain enzymolysis D.barbasco Chinese yam, and the saponin productive rate does not exceed direct acid-hydrolysis method level; The biological institute in Sichuan once carried out the research of pre-fermentation raising diosgenin productive rate and starch comprehensive utilization aspect, and propose that " Rhizome of Peltate Yam has kind can be influenced the enzyme that saponin is converted into sapogenin and exist; when the catalytic condition of suitable this kind of enzyme exists; its enzymic activity is strong, just can promote the raising of saponin productive rate in pre-fermenting process.Otherwise, after this kind of enzyme activity is damaged, though handle by pre-fermentation condition, all can not improve the saponin productive rate ", thereby, " without pre-fermentation, high temperature when zymamsis, high pressure steam material, have just played the effect of killing the saponin enzyme, so can not improve the saponin productive rate." that is the enzyme that produces during the fermentation of distiller's yeast, can not make saponin be degraded to secondary glycoside, thereby can not replace " from enzymic fermentation "; After 1 year, boiling at once after light industry institute in Shaanxi Province's pulverizes Rhizome of Peltate Yam, saccharification insert 1300 ^#Yeast carries out zymamsis, and per hundred jin of Chinese yams produce 96 degree 12.65 jin of alcohol (the rate of getting alcohol 27.3%), produce 0.78 jin of saponin (saponin content 0.9 to 1.5% in the raw material).

Can they only be conceived to utilize starch, improve the saponin yield to yeast fermentation and do not inquired into.The present invention is the disadvantage of radical cure " from enzymic fermentation ", several purebred black-koji moulds have been screened, carried out repeatedly fermenting experiment, cut-off connects total sapogenin that acid-hydrolysis method extracts then, after purebred black-koji mould fermentation, the total sapogenin that extracts through acid hydrolysis contrasts with the pre-saponin standard specimen of potato again, carry out tlc analysis, obtained two pairs of color spots.First couple of color spot Rf=0.45, in full accord with the Rf value of standard specimen, visible this color spot is exactly the color spot of diosgenin.Second pair color spot is little and look shallow, Rf=0.85, and this is the color spot of other sapogenin or derivatives thereofs of coexistence, does not see other obvious color spots in addition.

To ferment and the total sapogenin sample of unfermentable two classes, gradation is successively used ether and benzene wash-out after the absorption of alumina dry post, obtain two elution peaks, collects product respectively, measures infrared spectra.As a result, the infrared spectra of the ether eluate of this two classes sample is identical ν ^Film _MaxCm ^-1: 1057(C ₃-OH and △ ⁵), 986,926＜904, the 868(25R spiral shell stays alkane), contrasting decidable with Sadler's commodity spectrogram S-092 is diosgenin; Mass spectrum M/Z:414(M ⁺), the 139(base peak) 282,300, be diosgenin through library searching also decidable.The infrared spectra of the benzene eluate of this two classes sample is also identical, and they are same derivatives.

The above results shows, the change of black-koji mould fermentation not causing diosgenin molecular structure, and derivative is an inherent in the raw material, rather than produce in the fermentation.

Use without the general raw materials for production from enzymic fermentation, destroyed after the various enzymes that self exist through boiling, directly use the black-koji mould inoculation fermentation, extract diosgenin through acid hydrolysis again, its yield has improved more than 100% output all 4～5%.

In addition, it is raw material that the present invention adopts yam, through the black-koji mould one time fermentation, can produce two kinds of product-diosgenins and citric acid simultaneously on a cover production line.In the yam rhizome, except that containing various saponins and Mierocrystalline cellulose, also contain a large amount of starch, in enzymic fermentation, perhaps be decomposed into water and CO original ₂Run away, perhaps mould bad, perhaps drain with spent acid solution, both enabled from acid solution, further to utilize starch, also neutralization in advance just can transfer other products to through Secondary Fermentation again.The present invention then is, under the black-koji mould effect, only needs one time fermentation, Yi Bian make the starch hydrolysis and change citric acid into, Yi Bian make sugar chain part enzymolysis in the diosgenin molecule, becomes secondary glycoside and sugar.(shown in the accompanying drawing schema) by fermentation can extract citric acid respectively from fermented liquid, extract diosgenin from filter residue.Through the check of Hubei bureau of import ﹠ export commodities inspection, the diosgenin of extraction and citric acid all reach export standard.

Recommendable implementation condition is as follows:

1. 5～15 gram tapioca flours are put into the 250ml conical flask, add 50ml water, wrap bottleneck, with 1.15～1.5kg/cm ²Open steam sterilization, and gelatinization 20～40 minutes is chilled to below 40 ℃, seals bottleneck after the inoculation, is placed on the shaking table 30～40 ℃ of temperature controls, ferment 3～4 days, takes out and filters, to the content and the purity of filtrate mensuration citric acid; To filter residue 4NH ₂SO ₄Ordinary-pressure hydrolysis 2～4 hours filters and is washed till nearly neutrality, and the baking universe changes in the Kai Shi extractor, with the sherwood oil (or 120 of 60～90 ℃ of boiling ranges ^#Gasoline) lixiviate is 2～6 hours, concentrate then, and crystallization, drying is weighed and is surveyed fusing point, molten distance.

Also can adopt following processing condition.

2. at first the Chinese yam sheet is soaked, pump into behind the defibrination in the fermentor tank, make raw material: water=7～13: 50.Under agitation be warming up to 120～150 ℃ of sterilizations with steam, gelatinization 20～40 minutes is waited to be chilled to below 40 ℃ and can be inoculated, and 28～42 ℃ of temperature controls fermented 2～4 days.Stir speed (S.S.) is 120～180 rev/mins.Being close to zero with reducing sugar is fermentation termination.During fermentation ends, be warming up to 65～75 ℃ of boiling slips and squeeze into plate-and-frame filter press while hot and filter.Fermented liquid CaOO ₃In and after-filtration, affination, the white powder crystal be citrate of lime, can get citric acid or other salts through processing again.Filter residue adds acid size mixing after, at 0.8～4kg/cm ²Vapour pressure under, hydrolysis 2～6 hours is squeezed into the plate-and-frame filter press press filtration then and is washed till nearly neutrality, drops into extractor through air stream drying to water content＜5% o'clock, with 120 ^#The gasoline lixiviate is to clean.Be concentrated into gasoline at concentration tank: saponin=30～60: changed over to and to weigh after crystallizer crystallization, filtration, the drying and survey fusing point, molten distance at 1 o'clock.

Because black-koji mould fermentation is without any negative reaction, and by fermentation with hydrolysis after twice filtration washing, in the hydrolyzate pigment and other impurity all seldom, so constant product quality, fusing point, molten be standard up to standard apart from primary crystallization.

Here, provide three embodiment.

Example 1 is got 1 part of tapioca flour, adds 5 parts of water, seals bottleneck with gauze and kraft paper in the 250ml conical flask of packing into, with 1.2～1.5kg/cm ²Steam sterilizing 20 minutes is chilled to below 40 ℃, from the inclined-plane that refrigerator is preserved, gets B ₈₂₆Bacterial classification one ring is put into through adequately disinfected pure water, smashes with glass sphere and makes its suspension, gets suspension 1ml and injects conical flask by syringe, shaking table top fermentation 114 hours.Total reducing sugar be can get thus and sour rate 95.0%, citric acid purity 98.9%, diosgenin 4.48%, yield 88.4% changeed.

Example 2 is got 11 parts of tapioca flours, adds 50 parts of water, after the sterilization, inoculates B in conical flask ₈₂₈Melanomyces spore suspension 1ml fermentation 110 hours.Total reducing sugar be can get thus and sour rate 85.9%, citric acid purity 91.1%, diosgenin yield 4.75%, yield 93.3% changeed.

Example 3 is got 11 parts of tapioca flours, adds 50 parts of water, adds a small amount of Ye Huamei, according to inoculating B with method sterilization back ₈₂₈Black-koji mould suspension 1ml fermented 4 days.Total reducing sugar be can get and sour rate 90.7%, citric acid purity 71.1%, diosgenin yield 4.95%, yield 97.6% changeed.

Claims (3)

1, a kind of is the novel process of raw material coproduction diosgenin and citric acid with the yam that contains the pre-saponin of potato, it is characterized in that improving the saponin yield with the black-koji mould fermentation.

2, novel process as claimed in claim 1 is characterized in that remaining unchanged by the molecular structure of fermentation diosgenin.

3, novel process as claimed in claim 1 is characterized in that with a kind of raw material just can obtaining two products of diosgenin and citric acid simultaneously only through one time fermentation, also can appoint in the production and get one.

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