CN104357507B

CN104357507B - A kind of high concentration L sorbose fermentation manufacturing techniques

Info

Publication number: CN104357507B
Application number: CN201410561050.7A
Authority: CN
Inventors: 肖江锋; 黄建民; 冯丽萍; 杜尔凤; 陶海; 赵杜娟
Original assignee: DSM Jiangshan Pharmaceutical Jiangsu Co Ltd
Current assignee: DSM Jiangshan Pharmaceutical Jiangsu Co Ltd
Priority date: 2014-10-21
Filing date: 2014-10-21
Publication date: 2017-08-22
Anticipated expiration: 2034-10-21
Also published as: CN104357507A; WO2016062232A1

Abstract

The invention discloses the L sorbose high-efficiency fermenting production technologies in the case of a kind of high concentration, using raw black gluconobacter suboxydans as production bacterial strain, L sorboses are produced by fermenting substrate of D sorbierites, 0.1mg/ml~5mg/ml gelatin is added in kind of liquid culture medium formula, in the fermented and cultured any time of 0~16 hour, addition 0.1mg/ml~5mg/ml citric acids, 0.1mg/ml~5mg/ml itaconic acids are individually added or combined into zymotic fluid.Present invention improves raw black gluconobacter suboxydans cell growth state, kind of a liquid quality is improved；Allogenic material citric acid, itaconic acid are added in fermentation process, promote raw black gluconobacter suboxydans cell metabolism, so as to improve in fermentation process, L sorbs sugared content is all higher than 420mg/ml after L sorboses generating rate, fermentation, has reached the purpose for improving fermentation efficiency and shortening fermentation period.

Description

A kind of high concentration L- sorbose fermentation manufacturing techniques

Technical field

The invention belongs to ascorbic preparing technical field, more particularly to a kind of high concentration L- sorboses fermenting and producing work Skill.

Background technology

Vitamin C is a kind of necessary water soluble vitamin of human body, with inoxidizability, improves the work such as immunity of organisms With being widely used in the fields such as medicine, food, cosmetics.

It is production of vitamin C by D-glucitol fermentation generation L- sorboses in current industrial vitamin C production technology An indispensable step in process route is industrial general using raw black gluconobacter suboxydans (Gluconobacter Melagenus) as production bacterium, during fermenting and producing L- sorboses in the case of high concentration substrate, thalline life is still suffered from The technical problem of the restriction production such as long suppressed, fermentation period length, energy consumption height, therefore, improves original fermentation manufacturing technique, improves L- sorbose fermentation efficiencies are very important.

In order to solve the above problems, it is (public that our company develops a kind of processing method of raising L- sorbose fermentation productivities The number of opening is CN103114111A), raw black gluconobacter suboxydans are used for production bacterial strain the invention provides one kind, with D-glucitol For fermenting substrate produce L- sorboses, in the fermented and cultured any time of 0~12 hour, into zymotic fluid add 0.01%~ 0.05% reduced glutathione, the biosynthesis speed ratio prior art of L- sorboses can at most improve 31.2%；In fermentation Any time of culture 0~12 hour, 0.01%~0.05% casamino acid, the biology of L- sorboses are added into zymotic fluid Synthesis rate can improve 27.9% than prior art.L- sorbose fermentation periods can effectively be shortened by above technical scheme, carried High vitamin c fermenting production efficiency.Data below is also listed in the embodiment scheme of the patent：Fermented in embodiment 1 24 hours sorb sugared content 287.2mg/ml；Ferment 26 hours sorb sugared content 307.8mg/ml in embodiment 2；In embodiment 3 Ferment 24 hours sorb sugared content 270.6mg/ml；Fermentation in 25 hours measures sorb sugared content and is to terminal in embodiment 4 315.4g/L.In actual use, the maximum sorb sugared content only 315.4g/L obtained after program fermentation, still suffers from hair The problem of ferment concentration is low.

The content of the invention

The purpose of the present invention will solve above-mentioned technical problem can at most improve there is provided a kind of relatively control of generating rate, effectively Shorten a kind of high concentration L- sorbose fermentation manufacturing techniques of L- sorbose fermentation periods.

In order to realize above-mentioned purpose, this invention takes following technical solution：A kind of high concentration substrate condition Under L- sorbose high-efficiency fermenting production technologies, it is characterised in that：

Its processing step is as follows：

1st, strain：Raw black gluconobacter suboxydans；

2nd, culture medium prescription：

Shaking flask kind liquid culture medium (mg/ml)：D-glucitol 150, yeast extract 5, peptone 5, calcium carbonate 1.5；Glacial acetic acid is adjusted Save pH to 4.8~5.4,121 DEG C of autoclaving 25min；

Plant liquid and expand culture medium (mg/ml)：D-glucitol 150, yeast extract 3, peptone 3, calcium carbonate 1.5；Glacial acetic acid is adjusted Save pH to 4.8~5.4,121 DEG C of autoclaving 25min；

Ferment initial medium (mg/ml)：D-glucitol 450, yeast extract 1.5, peptone 1.5, calcium carbonate 1.5；Ice vinegar Acid for adjusting pH is to 4.8~5.4,121 DEG C of autoclaving 25min；

3rd, prepared by shaking flask kind liquid：

The black gluconobacter suboxydans of the life of picking in right amount are into kind of liquid culture medium from slant medium, shaking flask volume 750ml, Liquid amount 250ml, 32 ± 1 DEG C, shaking speed 230r/min, culture 14h is standby as kind of liquid, control kind liquid L- sorb sugared contents 80mg/ml~120mg/ml；

4th, two grades of kind liquid incubation control：

Inoculum concentration is 1%, 33 ± 1 DEG C of cultivation temperature, and culture 16h is standby as kind of liquid, control kind liquid L- sorb sugared contents 80mg/ml~120mg/ml；

5th, fermentation processes：

Initial pH value 4.8~5.4,34 ± 1 DEG C of cultivation temperature, 500~800r/min of mixing speed, fermented and cultured 0~ 16h any time, 0.1mg/ml~5mg/ml citric acids, 0.1mg/ml~5mg/ml itaconic acids are added into zymotic fluid, after Supervention ferment is to terminal；

The present invention adds 0.1mg/ml~5mg/ml gelatin in two grades of kind liquid culture medium formulas, ferments and trains in L- sorboses During supporting 0~16h, the generating rate for adding L- sorboses in 0.1mg/ml~5mg/ml citric acids, its zymotic fluid is relatively compareed most 19.82% can be improved more；During 0~16h of L- sorbose fermented and cultureds, 0.1mg/ml~5mg/ml itaconic acids are added, it is sent out The generating rate of L- sorboses, which is relatively compareed, in zymotic fluid can at most improve 24.56%；During 0~16h of L- sorbose fermented and cultureds, Add the generation speed of L- sorboses in 0.1mg/ml~5mg/ml citric acids, 0.1mg/ml~5mg/ml itaconic acids, its zymotic fluid Rate, which is relatively compareed, can at most improve 26.32%.

The present invention is by adding allogenic material gelatin in L- sorboses fermentation kind liquid incubation, improving Sheng Hei Portugals Grape saccharic acid bacilli-cell growth conditions, improve kind of a liquid quality；Allogenic material citric acid, itaconic acid are added in fermentation process, is promoted Raw black gluconobacter suboxydans cell metabolism is entered, so as to improve in fermentation process L- mountains after L- sorboses generating rate, fermentation Pears sugared content is all higher than 420mg/ml, has reached the purpose for improving fermentation efficiency and shortening fermentation period, popularization and application have good Good economic and social benefit.

Embodiment

The invention will be further described below, but not as limitation of the present invention：

Embodiment 1, adds 1.5mg/ml gelatin in former two grades of kind liquid culture mediums, 2.5mg/ is added in the fermentation medium Ml citric acid, gives birth to black two grades of kind liquid of gluconobacter suboxydans, by cultured two grades of kind liquid using the culture of 10L Fermentations In 7.5L, 100L Fermentations of the access containing 50L fermentation mediums, initial pH value 5.2,34 DEG C of cultivation temperature, stirring speed 650r/min is spent, to terminal, L- sorb sugared content 427.20mg/ml produce sugared speed 10.95mg/ml/h, L- sorb to fermentation 39h Sugared throughput rate improves 17.87% compared with control.

Embodiment 2, adds 2.5mg/ml gelatin in former two grades of kind liquid culture mediums, 2mg/ml is added in the fermentation medium Itaconic acid, black two grades of kind liquid of gluconobacter suboxydans are given birth to using the culture of 10L Fermentations, by cultured two grades of kind liquid In 7.5L, 100L Fermentations of the access containing 50L fermentation mediums, initial pH value 5.3,34 DEG C of cultivation temperature, stirring speed 650r/min is spent, to terminal, L- sorb sugared content 423.90mg/ml produce sugared speed 11.16mg/ml/h, L- sorb to fermentation 38h Sugared throughput rate improves 21.04% compared with control.

Embodiment 3, adds 4mg/ml gelatin in former two grades of kind liquid culture mediums, 0.5mg/ml is added in the fermentation medium Citric acid, 2.5mg/ml itaconic acid gives birth to black two grades of kind liquid of gluconobacter suboxydans using the culture of 10L Fermentations, will train In the two grades of kind liquid 7.5L supported, 100L Fermentations of the access containing 50L fermentation mediums, initial pH value 5.2, culture temperature 34 DEG C of degree, mixing speed 650r/min, to terminal, L- sorb sugared content 422.7mg/ml produce sugared speed 11.42mg/ to fermentation 37h Ml/h, L- sorbose throughput rate improve 24.40% compared with control.

Embodiment 4, is added 2.5mg/ml gelatin in former two grades of kind liquid culture mediums, is given birth to using the culture of 10L Fermentations Black two grades of kind liquid of gluconobacter suboxydans, by cultured two grades of kinds liquid 7.5L, 100L of the access containing 50L fermentation mediums is automatic In fermentation tank, initial pH value 5.3,34 DEG C of cultivation temperature, mixing speed 650r/min；Cultivate after 4h, added into zymotic fluid 1mg/ml citric acid, 0.5mg/ml itaconic acid, to terminal, L- sorb sugared content 420.6mg/ml produce sugared speed to fermentation 38h 11.07mg/ml/h, L- sorbose throughput rate improve 21.11% compared with control.

Obviously, above-described embodiment is only intended to clearly illustrate example of the present invention, and not to embodiment Limit.For those of ordinary skill in the field, other multi-forms can also be made on the basis of the above description Change or variation.There is no necessity and possibility to exhaust all the enbodiments.And thus amplify out it is apparent Change or change still in protection scope of the present invention among.

Claims

1. a kind of high concentration L- sorbose fermentation manufacturing techniques, it is characterised in that：It is zymophyte using raw black gluconobacter suboxydans Strain, adds gelatin, fermentation process produces L- sorboses by substrate of D-glucitol in kind of liquid culture medium formula.

2. a kind of high concentration L- sorbose fermentation manufacturing techniques according to claim 1, it is characterised in that：The gelatin adds Dosage is 0.1 mg/ml~5mg/ml.

3. a kind of high concentration L- sorbose fermentation manufacturing techniques according to claim 1 or claim 2, it is characterised in that in L- sorbs In sugared fermentation process, citric acid and/or itaconic acid are added into zymotic fluid.

4. a kind of high concentration L- sorbose fermentation manufacturing techniques according to claim 3, it is characterised in that：The citric acid Addition is 0.1mg/ml~5mg/ml.

5. a kind of high concentration L- sorbose fermentation manufacturing techniques according to claim 3, it is characterised in that：The itaconic acid Addition is 0.1mg/ml~5mg/ml.