CN102296102B - Control method for gluconate production by microbiological method - Google Patents
Control method for gluconate production by microbiological method Download PDFInfo
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Abstract
The invention discloses a control method for gluconate production by a microbiological method. A high-yield gluconic acid strain is adopted as an original strain, which is moved into a fermentation tank for fermentation production of gluconate after seed culture. During fermentation, according to an on-line respiratory quotient (RQ), the adding speed and adding amount of glucose can be controlled in real time. The control method of the invention can make the glucose consumption rate reach 12g/L/h, and the conversion rate of production in the fermentation tank reach over 110%, thus realizing automatic flow feeding and substantially reducing the production cost.
Description
Technical field
The present invention relates to bioengineering field, specifically is the control method that a kind of microbial method is produced gluconate.
Background technology
Glucono-is as leavening agent, peptizer, intercalating agent, acidic flavoring agent and be widely used in industries such as food, medicine, chemical industry, water treatment, building; The glucono-salt (as with sodium, calcium, zinc, the synthetic gluconate that makes of MOX such as ferrous) can be used as foodstuff additive and accessory substance.
The production of glucono-mainly contains methods such as microbe fermentation method, electrolytic process and catalyzed oxidation.Because microbial method, mild condition is energy-conservationly obviously extensively adopted, and wherein microbial fermentation has fungi fermentation and fermentation using bacteria technology again.Since Boot Shandong (Boutroux) utilizes the oxygenizement of mikrobe can glucose oxidase be become glucono-first; Glucono-and its esters zymotechnique there has been tangible progress; As block beautiful (Currie) etc. and utilize mould or aspergillus to set up modern aeration-agitation liquid fermentation technology, make the transformation efficiency of glucose reach 90%.Blom etc. have invented interpolation NaOH, have kept the method production Sunmorl N 60S of pH value below 6.5, and inversion rate of glucose reaches more than 95%; China's ecliptic shakes etc. are the black mold seed liquor of 30% fermention medium inoculation 10% with glucose content, 6 0～6.5 residual sugar can be reduced to below 1 g/L through stream hydro-oxidation sodium solution control pH value.In addition, there have report to find that through medium optimization pH value, temperature, fermentor tank rotating speed, substrate glucose concn influence inversion rate of glucose to be bigger, obtains optimal processing parameter.Liu Zhong converges and waits the special-purpose pH temperature intelligent system that designs to industrial bulk fermentation, has obtained fermentation level preferably.In addition; The short stalk of A Disitaxiadisi employings such as (Anastassiadis) in 2002 mould (Aureobasidium pullulans) has been set up the continuous glucono-production technology of resting cell and immobilized cell; Concentration can reach 26%, and the highest generating rate is 19g/L/h.
Therefore; When glucono-and salt (like Sunmorl N 60S, calglucon etc.) fermentative prodn thereof, mainly adopt mold fermentation to carry out; Its fermentation control control method mainly is methods such as control pH, temperature, fermentor tank rotating speed, dissolved oxygen concentration (DO), remaining sugar concentration; Yet these methods all fail to observe the microorganism growth period of glucono-and salt fermentation thereof, more fail to use online RQ and realize fermenting process controlled on-line feed supplement strategy.
In sum; At present but this area lacks the method for glucono-and salt fermenting process line closed loop control flow feeding thereof, therefore press for exploitation a kind of online, accurately measure the method for mikrobe physiological metabolism respiratory quotient and control auto-feeding substrate glucose thereof.
Summary of the invention
The object of the invention just provide a kind of microbial method produce gluconate online, accurately measure the method for mikrobe physiological metabolism respiratory quotient and control auto-feeding substrate Glucose Liquid thereof.For realizing above-mentioned purpose, the present invention has adopted following technical scheme:
Microbial method is produced gluconate, the first step, and seed culture adds bacterial classification in the seed fermentation jar and substratum carries out seed culture, and said bacterial classification is black mold, Rhizopus oryzae, mould; Second step, fermentation, Glucose Liquid with 30% and nutrition agent add fermentor tank and sterilization, and cultured seed is inserted in the fermentor tank in 10% ratio, and 50%～60% of total add-on to fermentor tank working volume is installed in line analysis fermentation tail gas CO on the fermentor tank
2And O
2The exhaust analyzer of concentration or process mass spectrograph have also been installed the mass flowmeter of entering and exhausted air and the true microorganism cells volume of culture transmitter of fermentor tank simultaneously; The 3rd step, feed supplement, the aforementioned second step operation fermentation 17～19 hours; Go into 500g/L Glucose Liquid and nutrition agent according to carrying out on-line automatic benefit, be specially, add 500g/L Glucose Liquid and nutrition agent when RQ is lower than 0.05 ± 0.01 in the respiratory quotient (RQ) of line computation; The fermentor tank cubic capacity is made as V; Unit is L, and maximum liquid amount is controlled at the fermentor tank cubic capacity below 80%, and the speed S of adding is:
S=
(L/h)
Stop to add 500g/L Glucose Liquid and nutrition agent when RQ is higher than 0.05 ± 0.01, RQ is maintained between 0.05 ± 0.01, thereby can make the gluconic acid fermentation process steady, realize that maximum yield reaches more than the 12g/L/h.
Control method of the present invention is controlled Glucose Liquid in real time and is added speed and addition according to respiratory quotient (RQ) numerical value, has realized direct, real-time, the accurate control of the fermenting process of glucono-.
Embodiment
Further specify the present invention below in conjunction with embodiment.
Embodiment 1:
The first step, seed culture: add glucose 2.5Kg, steeping water 75g, potassium primary phosphate 6.4g, urea 2g, skimmer 2ml in the 15L culture tank; Be settled to 7.5L; Sterilized 15 minutes for 120 ℃, reduce to 38 ℃ then, add 250ml eggplant bottle bacteria suspension; Control dissolved oxygen (DO) >=30 is cultivated 15h and is got seed liquor.
Second step, fermentation: add glucose 4.2KG, sal epsom 2.8g, potassium primary phosphate 0.28g, urea 1.4g, skimmer 1ml in the 50L fermentor tank; Be settled to 13L; Sterilized 15 minutes for 120 ℃, reduce to 38 ℃ then, the inoculum size with 10% inserts cultured seed liquid.Control dissolved oxygen (DO) >=30,38 ℃ of temperature, the lime carbonate with 20% is regulated potential of hydrogen PH 5.5, begins fermentation.
The 3rd step, feed supplement:
(1). batching: add Glucose Liquid 14Kg, steeping water 36g, sal epsom 5.6g in the 30L feed supplement jar, urea 2.8g, skimmer 1ml is settled to 20L, and 120 ℃ of sterilizations 15 minutes are reduced to 38 ℃ then, and it is aseptic that to change serum bottle over to subsequent use.
(2). feed supplement control method: in the second step operation; Ferment about 18 hours according to carrying out on-line automatic 500g/L Glucose Liquid and the nutrition agent of adding in the feed supplement jar in the respiratory quotient (RQ) of line computation; Be specially, RQ=0.05 ± 0.01 is set, add 500g/L Glucose Liquid and nutrition agent when RQ is lower than 0.05 ± 0.01 stream; Stop stream and add 500g/L Glucose Liquid and nutrition agent when RQ is higher than 0.05 ± 0.01; RQ is maintained between 0.05 ± 0.01, thereby can make the gluconic acid fermentation process steady, realize that maximum yield reaches more than the 12g/L/h.Be specially: when fermentation during 11h, RQ=0.049, peristaltic pump is started working automatically, adds 500g/L Glucose Liquid and nutrition agent, and along with the benefit of Glucose Liquid is gone into, RQ paces up and down about 0.05 at 11-13h always, and the feed supplement pump is also opened start-stop and is stopped along with the variation of RQ numerical value.But RQ begins to reduce behind the 13h, and RQ is stabilized in 0.03 ± 0.005 after the 15h, and glucose is also added according to certain speed always.When the fermented liquid TV reaches fermentor tank volumetrical 80%, stop feed supplement, continuing to ferment to dissolved oxygen (DO) reaches at 100 o'clock, fermentation ends.
The 4th step, filtration: Plate Filtration.
The 5th step, decolouring: activated carbon decolorizing.
The 6th step, crystallization: 120-125 ℃ are concentrated into 70% above content, crystallisation by cooling.
The 7th step, separation, drying: separating machine is with crystal and liquid separation, and fluidised bed drying gets the calglucon finished product then.
The 8th step, packing.
Embodiment 2:
The first step, seed culture: add glucose 2.5Kg, steeping water 75g, potassium primary phosphate 6.4g, urea 2g, skimmer 2ml in the 15L culture tank; Be settled to 7.5L; Sterilized 15 minutes for 120 ℃, reduce to 38 ℃ then, add 250ml eggplant bottle bacteria suspension; Control dissolved oxygen (DO) >=30 is cultivated 15h and is got seed liquor.
Second step, fermentation: add glucose 4.2KG, sal epsom 2.8g, potassium primary phosphate 0.28g, urea 1.4g, skimmer 1ml in the 50L fermentor tank; Be settled to 13L; Sterilized 15 minutes for 120 ℃, reduce to 38 ℃ then, the inoculum size with 10% inserts cultured seed liquid.Control dissolved oxygen (DO) >=30,38 ℃ of temperature, the sodium hydroxide with 32% is regulated potential of hydrogen PH 5.5, begins fermentation.
The 3rd step, feed supplement:
(1). batching: add Glucose Liquid 14Kg, steeping water 36g, sal epsom 5.6g in the 30L feed supplement jar, urea 2.8g, skimmer 1ml is settled to 20L, and 120 ℃ of sterilizations 15 minutes are reduced to 38 ℃ then, and it is aseptic that to change serum bottle over to subsequent use.
(2). feed supplement control method: in the second step operation; Ferment about 18 hours according to carrying out on-line automatic 500g/L Glucose Liquid and the nutrition agent of adding in the feed supplement jar in the respiratory quotient (RQ) of line computation; Be specially, RQ=0.05 ± 0.01 is set, add 500g/L Glucose Liquid and nutrition agent when RQ is lower than 0.05 ± 0.01 stream; Stop stream and add 500g/L Glucose Liquid and nutrition agent when RQ is higher than 0.05 ± 0.01; RQ is maintained between 0.05 ± 0.01, thereby can make the gluconic acid fermentation process steady, realize that maximum yield reaches more than the 12g/L/h.Be specially: when fermentation during 11h, RQ=0.049, peristaltic pump is started working automatically, adds 500g/L Glucose Liquid and nutrition agent, and along with the benefit of Glucose Liquid is gone into, RQ paces up and down about 0.05 at 11-13h always, and the feed supplement pump is also opened start-stop and is stopped along with the variation of RQ numerical value.But RQ begins to reduce behind the 13h, and RQ is stabilized in 0.03 ± 0.005 after the 15h, and glucose is also added according to certain speed always.When the fermented liquid TV reaches fermentor tank volumetrical 80%, stop feed supplement, continuing to ferment to dissolved oxygen (DO) reaches at 100 o'clock, fermentation ends.
The 4th step, filtration: Plate Filtration.
The 5th step, decolouring: activated carbon decolorizing.
The 6th step, crystallization: 120-125 ℃ are concentrated into 70% above content, crystallisation by cooling.
The 7th step, separation, drying: separating machine is with crystal and liquid separation, and fluidised bed drying gets the Sunmorl N 60S finished product then.
The 8th step, packing.
Claims (1)
1. a microbial method is produced the control method of gluconate, it is characterized in that: the first step, in the seed fermentation jar, add bacterial classification and substratum carries out seed culture, and said bacterial classification is black mold, Rhizopus oryzae or mould; In second step, inoculation fermentation, Glucose Liquid with 30% and nutrition agent add fermentor tank and sterilization, add 10% seed culture fluid, 50%～60% of total add-on to fermentor tank working volume; The 3rd step, feed supplement, the aforementioned second step operation fermentation 17～19 hours; According to carrying out on-line automatic 500g/L Glucose Liquid and the nutrition agent added in the respiratory quotient (RQ) of line computation, be specially, when being lower than 0.04, RQ adds 500g/L Glucose Liquid and nutrition agent; The fermentor tank cubic capacity is made as V; Unit is L, and maximum liquid amount is controlled at the fermentor tank cubic capacity below 80%, and the speed S of adding is:
S=?
(L/h)
Stop to add 500g/L Glucose Liquid and nutrition agent when RQ is higher than 0.06, RQ is maintained between 0.05 ± 0.01.
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| CN103352089A (en) * | 2013-06-17 | 2013-10-16 | 山东福洋生物科技有限公司 | Control method for microbial fermentation production of gluconate |
| CN103352090A (en) * | 2013-06-18 | 2013-10-16 | 山东福洋生物科技有限公司 | Control method for efficient production of calcium gluconate by microbial fermentation |
| CN104805138A (en) * | 2015-04-07 | 2015-07-29 | 山东西王糖业有限公司 | Fermentation method of high-dry-matter sodium gluconate |
| CN105255963B (en) * | 2015-11-11 | 2019-03-01 | 山东福洋生物科技有限公司 | Curdlan high-efficiency fermenting control method |
| CN106349055B (en) * | 2016-08-24 | 2017-12-01 | 山东福洋生物科技有限公司 | A kind of new separating and extracting process of gluconic acid mother liquid of sodium |
| CN107881204A (en) * | 2017-11-24 | 2018-04-06 | 山东福洋生物科技有限公司 | A kind of method of fermentation of Aspergillus niger production sodium gluconate |
| CN111334538B (en) * | 2020-03-20 | 2023-04-11 | 鄂州职业大学 | Method for producing gluconic acid by strengthening penicillium funiculosum fermentation glucose |
| CN113249413A (en) * | 2020-07-08 | 2021-08-13 | 山东福洋生物科技股份有限公司 | Method for producing sodium gluconate by recycling aspergillus niger mycelia |
| CN114196713A (en) * | 2021-11-25 | 2022-03-18 | 山东润德生物科技有限公司 | Method for reducing carbon dioxide emission in glucosamine fermentation process |
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| CN1188152A (en) * | 1997-12-17 | 1998-07-22 | 中山大学 | Producing of magnesium (manganese) gluconate by aspergillus fermentation glucose |
| CN102071224A (en) * | 2009-11-25 | 2011-05-25 | 华东理工大学 | Method for producing sorbitol and gluconate |
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| CN1188152A (en) * | 1997-12-17 | 1998-07-22 | 中山大学 | Producing of magnesium (manganese) gluconate by aspergillus fermentation glucose |
| CN102071224A (en) * | 2009-11-25 | 2011-05-25 | 华东理工大学 | Method for producing sorbitol and gluconate |
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| Yuko Ikeda et al..Bioconversion of waste office paper to gluconic acid in a turbine blade reactor by the filamentous fungus Aspergillus niger.《Bioresource Technology》.2006,第97卷(第8期),1030-1035. * |
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Address after: The economic development zone of Shandong city in Dezhou province 253100 County Road East Petroleum Company Fuyang plain Patentee after: Shandong Fu Yang biological Polytron Technologies Inc Address before: The economic development zone of Shandong city in Dezhou province 253100 County Road East Petroleum Company Fuyang plain Patentee before: Shandong Fuyang Biotechnology Co., Ltd. |