CN1164763C - Biological fermentation culture medium substrate and its preparation method - Google Patents
Biological fermentation culture medium substrate and its preparation method Download PDFInfo
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- CN1164763C CN1164763C CNB011061731A CN01106173A CN1164763C CN 1164763 C CN1164763 C CN 1164763C CN B011061731 A CNB011061731 A CN B011061731A CN 01106173 A CN01106173 A CN 01106173A CN 1164763 C CN1164763 C CN 1164763C
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Abstract
The present invention relates to a substitute of the yeast cream and the yeast powder of a biological fermentation culture medium and a preparation method thereof by utilizing penicillin bacteria slag. The preparation method comprises the process steps of strain preparation, pretreatment, the culture in a fermentation tank, separation, extraction, etc. The present invention has the advantages of low production cost, reasonable design in production processes, simplicity, high speed, reliability, easy operation, and stable product quality; the substitute is suitable for large-scale commercial product; the substitute surpasses normal yeast cream and normal yeast powder through the experimental result of application; the substitute solves the environmental pollution problem caused by waste slag in the fermentation production of penicillin; the substitute provides a new path for the clean production of penicillin fermentation industry.
Description
The present invention relates to a kind of biological fermentation culture medium substrate and preparation method thereof, a kind of specifically penicillin mushroom dregs of utilizing prepares yeast extract paste in the biological fermentation culture medium, yeast powder surrogate and preparation method thereof.
Penicillin mushroom dregs is the main waste material in China's antibiotic fermentation industrial production, wherein contain a large amount of tropinas, account for more than 40% of dry weight, abundant VITAMIN, nutritive substances such as somatomedin and substratum residue, also and depositing the various meta-bolitess of microorganism, the downstream processing additive, utilization for penicillin mushroom dregs, domestic have minority producer to utilize the exsiccant method to produce the feedstuff protein surrogate, this simple physics drying means, in fact can't remove above poisonous substance fully, this is the still unsolved technical problem underlying of present penicillin mushroom dregs development and use, so range of application is greatly limited.The deep development of penicillin mushroom dregs is in the blank stage especially, so up to the present, a large amount of penicillin mushroom dregs are still abandoned as waste, have not only wasted a large amount of precious resources, and severe contamination environment.
The new purposes that the objective of the invention is to the deep development penicillin mushroom dregs is carried out detoxification to penicillin mushroom dregs and is prepared the product innovation that replaces yeast extract paste, yeast powder with new processing method, for the cleaner production of penicillin fermentation industry provides technical support.
The object of the present invention is achieved like this:
One, the raw material for preparing yeast extract paste in the biological fermentation culture medium, yeast powder surrogate with penicillin mushroom dregs
Major ingredient: penicillin mushroom dregs, the bright slag of dried slag all can.
Auxiliary material: glucose, sucrose, maltose, molasses, starch, dextrin, technical-grade hydrochloric acid, sulfuric acid, acetic acid, technical grade sodium hydroxide, potassium hydroxide, yellow soda ash, sodium bicarbonate, unslaked lime, ammonia.
Two, preparation method
1, the preparation of bacterial classification
The bacterial classification that uses is as forming in cereuisiae fermentum (S.cerevisiae Hansen) No.2.1190 and the compound cultivation liquid medium within of rhodopseudomonas (Rhodopseudomonas), two kinds of bacterium all derive from Heilongjiang Province using microbe institute, and medium component is: glucose 3%, ammonium sulfate 0.5%, lime carbonate 0.003%, Nacl 0.02%, yeast extract paste 0.2%; Culture condition: 30 ℃ ± 1 ℃, intermittently ventilation, air flow 1: 0.5-1: 0.8vvm, mixing speed 150rpm, culture cycle 2-3 days, mature seed liquid was PH5-5.5.
2, pre-treatment: penicillin mushroom dregs is added acid solution, and example hydrochloric acid liquid or other acid solution are transferred PH2-4, handle 4-8 hour for 60 ℃-80 ℃.Be neutralized to PH6.0-6.5 with alkali lye such as sodium hydroxide solution or other alkali lye.
3, fermentor cultivation: in fermentor tank, add 10% penicillin mushroom dregs, 1-5% glucose or other glucide, the water of 85-89% was sterilized 30 minutes for 121 ℃, when treating that temperature is reduced to 30 ℃ ± 2 ℃, insert the 1-2% seed liquor, at 30 ℃ ± 2 ℃, intermittently ventilation, air flow 10.4-10.8, mixing speed 150rpm cultivates and put a jar extraction in 4-5 days; Above-mentioned fermented liquid carries out solid-liquid separation after putting jar, collect filtrate, transfer PH6.5-7.0 with alkali lye as 20% sodium hydroxide or other alkali lye, left standstill 5-6 hour, collect supernatant liquor and change 121 ℃ of sterilizations of concentration tank 30 minutes over to, be cooled to 60 ℃ of vacuum concentration to proportion (60 ℃ of heat are surveyed) 1.30 then and promptly get the yeast extract paste surrogate.
The yeast extract paste surrogate is a tawny, and the thickness paste is slightly stuck with paste fragrance, moisture≤30%, total nitrogen 〉=6.0%, chlorine root≤5%, arsenic≤1mg/g, residue on ignition≤12%.
The preparation of yeast powder surrogate is in above-mentioned preparation process, fermented liquid is put jar after, transfer PH to 6.5, directly spraying drying.
The yeast powder surrogate is the tawny powder, slightly sticks with paste fragrance, protein 〉=40% wherein, molten phosphorus 4000-5000r/g, moisture 10%.
The present invention utilizes penicillin mushroom dregs to prepare yeast extract paste, the yeast powder surrogate is a kind of novelty exploitation to industrial residue, its low production cost, because main raw material is to replace conventional yeast powder with penicillin mushroom dregs, only this item main material cost just can reduce about 5000 yuan/ton, production technique is reasonable in design simultaneously, simple and direct reliable, processing ease, constant product quality, be fit to large-scale industrial production, the effect of this kind product innovation in biological fermentation culture medium is good, and experimental result has been better than conventional yeast extract paste and yeast powder, so have promotional value, the more important thing is during having solved penicillin fermentation produces waste residue to the pollution problem of environment, for the cleaning of penicillin fermentation industry produces the new approach started.
Fig. 1 is for the present invention produces yeast extract paste surrogate process flow sheet;
Fig. 2 is for the present invention produces yeast powder surrogate process flow sheet.
Embodiment 1
The preparation of yeast extract paste surrogate
Get 1 kilogram of dried penicillin mushroom dregs, add 6N technical hydrochloric acid 3-4Kg and be acidified to PH3-4, handled 4-8 hour for 60-80 ℃, transfer PH6.0-6.5 with 40% industrial sodium hydroxide, add 100 gram glucose, the constant volume fermentation volume is 10 kilograms, sterilized 30 minutes for 121 ℃, be cooled to about 30 ℃, insert the seed liquor of 1-2%, in 30 ℃ ± 2 ℃, intermittently ventilation, air flow 1: 0.5-1: 0.8vvm, mixing speed 150rpm, cultivated 4-5 days, blowing extracts.The gained fermented liquid is carried out solid-liquid separation, collect filtrate, transfer PH to 6.0-7.0 with 20% sodium hydroxide solution, left standstill 5-6 hour, collect supernatant liquor in 121 ℃ of sterilizations 30 minutes, under 60 ℃, be evaporated to then proportion (60 ℃ heat survey) 1.30 the yeast extract paste surrogate, yield about about 50%.
Embodiment 2
The preparation of yeast powder surrogate
In embodiment 1, behind the fermented liquid blowing, behind the adjusting PH to 6.5, directly spraying drying promptly gets the yeast powder surrogate, yield 90-100%.
Application Example 1
The yeast extract paste surrogate of the present invention's production is used for the shake flask fermentation test of Avrmectin (AVM), its operating process is 1. medium preparation: get starch 7%, analysis for soybean powder 0.8%, yeast extract paste 0.25%, (the test group yeast extract paste is the yeast extract paste surrogate with the penicillin mushroom dregs preparation, control group is conventional yeast extract paste), yeast powder 0.4%, sal epsom 0.02%, dipotassium hydrogen phosphate 0.05%, lime carbonate 0.002%, Yi Shui complement to 100%, transfer PH7.0, the triangular flask packing, loading amount is 100ml/500ml, sterilizes 30 minutes for 121 ℃.2. inoculation: treat that substratum is cooled to about 30 ℃, insert 1% bacterial classification.3. culture condition: postvaccinal fermentation flask is put on the bottle swingging machine, rotating speed 180rpm, 28 ℃ ± 1 ℃ cultivation of temperature is cultivated 9 bottles all over the world, with the high-pressure liquid chromatography chemical titer.Through the test more than 5 times, respectively 10 bottles of test group and contrasts are set up in each test, and test-results is 10 bottles of sample mean numbers, and result's yeast extract paste surrogate of the present invention improves 7%-14% than conventional yeast extract paste chemical titer, sees Table 1
Table 1:
| The test group sequence number | 1 | 2 | 3 | 4 | 5 |
| The test group μ g/ml that tires | 900 | 1068.7 | 1001.3 | 835 | 856 |
| The control group μ g/ml that tires | 786.7 | 996.3 | 900 | 730 | 786.7 |
| Test group is tired than control group and is improved % | 14.4 | 7.3 | 11.3 | 14.4 | 8.8 |
Application Example 2
The yeast powder surrogate of the present invention's production is used for the shake flask fermentation test of Avrmectin (AVM), with conventional yeast powder is control group, its operating process is its medium preparation: get starch 7%, analysis for soybean powder 0.8%, yeast powder 0.7%, sal epsom 0.02%, dipotassium hydrogen phosphate 0.05%, lime carbonate 0.002%, Yi Shui and complement to 100%, transfer PH7.0, the triangular flask packing, loading amount is 100ml/500ml, sterilizes 30 minutes for 121 ℃.2. inoculation: treat that substratum is cooled to about 30 ℃, insert 1% bacterial classification.3. culture condition: postvaccinal fermentation flask is put on the bottle swingging machine, rotating speed 180rpm, 28 ℃ ± 1 ℃ of temperature is cultivated 9 bottles all over the world, with the high-pressure liquid chromatography chemical titer.Through the test more than 5 times, respectively 10 bottles of test group and contrasts are set up in each test, and test-results is 10 bottles of mean numbers, and result's yeast powder surrogate of the present invention is roughly more suitable than conventional yeast powder chemical titer, sees Table 2
Table 2
| The test group sequence number | 1 | 2 | 3 | 4 | 5 |
| The test group μ g/ml that tires | 821.3 | 805.6 | 806.9 | 926.3 | 797.5 |
| The control group μ g/ml that tires | 810.9 | 751.3 | 810.9 | 975 | 759.5 |
| Test group is tired than control group and is improved % | 1.3 | 7.2 | -0.5 | -5.3 | 5.0 |
Application Example 3
The yeast powder surrogate of the present invention's preparation is used for the shake flask fermentation test of terramycin, with conventional yeast powder is control group, specific as follows: substratum is formed: starch 10%, dextrin 5%, soybean cake powder 2%, yeast powder 0.2%, lime carbonate 1.2%, ammonium sulfate 1.2%, sodium-chlor 0.4%, dipotassium hydrogen phosphate 0.015%, complement to 100% with water, loading amount is 40ml/500ml, every bottle adds soya-bean oil 1ml, sterilized 30 minutes for 121 ℃, inoculum size is the 5ml/ bottle, culture condition: shaking speed 200-220rpm, 30 ℃ of temperature, cultivate 8 world shaking tables, with 581 its chemical titers of type spectrophotometric instrumentation.Through 3 tests, respectively 10 bottles of test group and control groups are set up in each test, and its result is 10 bottles of mean numbers.Test-results shows that yeast powder surrogate of the present invention improves more than 7% than the average chemical titer of conventional yeast powder, sees Table 3.
Table 3
| The test group sequence number | 1 | 2 | 3 |
| The test group μ g/ml that tires | 32100 | 30210 | 28900 |
| The control group μ g/ml that tires | 28200 | 27500 | 26900 |
| Test group is tired than control group and is improved % | 13.8 | 9.8 | 7.4 |
Claims (9)
1, a kind of preparation method of fermention medium surrogate is characterized in that, this method may further comprise the steps:
(1) preparation of bacterial classification:
The bacterial classification of using is cereuisiae fermentum (S.CerevisiaeHansen) No.2.1190 and rhodopseudomonas (Rhodopseudomonas), in the compound cultivation liquid medium within of above two kinds of bacterium, medium component: glucose is about 3%, ammonium sulfate is about 0.5%, lime carbonate is about 0.003%, NaCl is about 0.02%, yeast extract paste about 0.2%, culture condition: 30 ℃ ± 1 ℃, intermittently ventilation, air flow 1: 0.5-1: 0.8vvm, mixing speed 150rpm, culture cycle 2-3 days;
(2) with the penicillin mushroom dregs be raw material, at first carry out pre-treatment: regulate PH3-4 with acid solution, handle for 60 ℃-80 ℃ and use in the alkali lye after 4-8 hour and PH6-6.5;
(3) the bacterium slag after the pre-treatment is dropped into fermentor tank, add-on is about 10% of fermentation volume, the carbon source that adds 1-5% again, water 85-89% sterilized 30 minutes for 121 ℃, to be cooled during to 30 ℃ ± 2 ℃, insert the seed liquor of 1%-2%,, stir 150rpm in 30 ℃ ± 2 ℃, intermittently ventilate 1: 0.5-1: 0.8vvm, cultivate after 4-5 days and fermented liquid to be put a jar extraction.
2, the prepared fermention medium surrogate of preparation method according to claim 1.
3, the described method of claim 1, when described culture medium substrate is the yeast powder surrogate, this method also comprise with described fermented liquid pH transfer to 6.5 the back convection dryings.
4, the described method of claim 1, when described culture medium substrate is the yeast extract paste surrogate, this method also comprises carries out solid-liquid separation with described fermented liquid, collect filtrate, transfer PH6-7, left standstill 5-6 hour, collect supernatant liquor and go into concentration tank, sterilized 30 minutes for 121 ℃, 60 ℃ of vacuum concentration to 60 ℃ proportions that heat is surveyed are 1.30.
5, the described fermention medium surrogate of claim 2 is characterized in that:
(1) the yeast extract paste surrogate is a tawny, and the thickness paste is slightly stuck with paste fragrance, moisture content≤30%, total nitrogen 〉=6.0%, chlorine root≤5%, arsenic≤1mg/g, residue on ignition≤12%;
(2) the yeast powder surrogate is the tawny powder, slightly sticks with paste fragrance, protein 〉=40% wherein, molten phosphorus 4000-5000r/g, moisture 10%.
6, preparation method according to claim 1 is characterized in that, wherein said acid solution is meant the aqueous solution of technical-grade hydrochloric acid, sulfuric acid, acetic acid.
7, preparation method according to claim 1 is characterized in that, wherein said alkali lye is meant the aqueous solution of technical grade sodium hydroxide, potassium hydroxide, yellow soda ash, sodium bicarbonate, ammonia.
8, preparation method according to claim 1 is characterized in that: wherein said carbon source is meant glucose, sucrose, maltose, honey, starch, dextrin.
9, penicillin mushroom dregs is used to make the purposes of the described culture medium substrate of claim 2.
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| CNB011061731A CN1164763C (en) | 2001-02-20 | 2001-02-20 | Biological fermentation culture medium substrate and its preparation method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101935682B (en) * | 2010-08-20 | 2012-07-18 | 石药集团中诺药业（石家庄）有限公司 | Application method for cephalosporin C bacterium residue |
| CN102286413B (en) * | 2011-09-03 | 2012-12-19 | 福建农林大学 | Preparation method of liquid fermentation medium for bacillus thuringiensis |
| CN102409073A (en) * | 2011-10-18 | 2012-04-11 | 江瀚生物科技（上海）有限公司 | Method for producing antioxidant of microbial food |
| CN103861447B (en) * | 2012-12-14 | 2015-09-30 | 哈尔滨科艺生物环保科技有限责任公司 | A kind of cultivation place cleanser and preparation method thereof |
| CN104223318A (en) * | 2014-08-25 | 2014-12-24 | 南通昊友食品添加剂有限公司 | Production method of microorganism food antioxidant |
| CN112725385B (en) * | 2019-10-28 | 2022-09-09 | 中国石油化工股份有限公司 | Method for preparing long-chain dicarboxylic acid by fermentation |
| CN114292755B (en) * | 2021-12-06 | 2024-01-26 | 内蒙古常盛制药有限公司 | Method for recycling penicillin fermentation from antibiotic fungus dreg lysate |
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