CN101402914B - Application of bagasse as immobilized material in immobilized fiber bed reactor for producing organic acid by fermentation - Google Patents
Application of bagasse as immobilized material in immobilized fiber bed reactor for producing organic acid by fermentation Download PDFInfo
- Publication number
- CN101402914B CN101402914B CN200810234514.8A CN200810234514A CN101402914B CN 101402914 B CN101402914 B CN 101402914B CN 200810234514 A CN200810234514 A CN 200810234514A CN 101402914 B CN101402914 B CN 101402914B
- Authority
- CN
- China
- Prior art keywords
- reactor
- immobilization
- bagasse
- feed supplement
- acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 239000000835 fiber Substances 0.000 title claims abstract description 102
- 241000609240 Ambelania acida Species 0.000 title claims abstract description 65
- 239000010905 bagasse Substances 0.000 title claims abstract description 65
- 238000000855 fermentation Methods 0.000 title claims abstract description 65
- 230000004151 fermentation Effects 0.000 title claims abstract description 65
- 150000007524 organic acids Chemical class 0.000 title claims abstract description 45
- 239000000463 material Substances 0.000 title claims abstract description 41
- 238000004519 manufacturing process Methods 0.000 claims abstract description 72
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 claims description 116
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 claims description 62
- 235000019260 propionic acid Nutrition 0.000 claims description 57
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 claims description 57
- 239000006052 feed supplement Substances 0.000 claims description 56
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 40
- 239000003513 alkali Substances 0.000 claims description 32
- 239000002253 acid Substances 0.000 claims description 29
- 239000004310 lactic acid Substances 0.000 claims description 20
- 235000014655 lactic acid Nutrition 0.000 claims description 20
- 238000003756 stirring Methods 0.000 claims description 19
- 239000000523 sample Substances 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 238000010298 pulverizing process Methods 0.000 claims description 6
- 210000004027 cell Anatomy 0.000 abstract description 27
- 238000000034 method Methods 0.000 abstract description 26
- 210000001822 immobilized cell Anatomy 0.000 abstract description 3
- 239000002699 waste material Substances 0.000 abstract description 3
- 230000007423 decrease Effects 0.000 abstract description 2
- 241000196324 Embryophyta Species 0.000 abstract 1
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Inorganic materials [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 87
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 79
- 239000008103 glucose Substances 0.000 description 79
- 229920000742 Cotton Polymers 0.000 description 29
- 230000001580 bacterial effect Effects 0.000 description 28
- 235000013311 vegetables Nutrition 0.000 description 28
- 239000002609 medium Substances 0.000 description 20
- 229960000448 lactic acid Drugs 0.000 description 19
- JVTAAEKCZFNVCJ-REOHCLBHSA-N L-lactic acid Chemical compound C[C@H](O)C(O)=O JVTAAEKCZFNVCJ-REOHCLBHSA-N 0.000 description 18
- 239000000243 solution Substances 0.000 description 17
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 16
- 239000001963 growth medium Substances 0.000 description 15
- 238000011218 seed culture Methods 0.000 description 15
- 239000007789 gas Substances 0.000 description 14
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 10
- 239000000499 gel Substances 0.000 description 10
- 239000002054 inoculum Substances 0.000 description 10
- 239000011261 inert gas Substances 0.000 description 8
- 239000001384 succinic acid Substances 0.000 description 8
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 7
- 241000894006 Bacteria Species 0.000 description 7
- 241000218588 Lactobacillus rhamnosus Species 0.000 description 7
- 239000001888 Peptone Substances 0.000 description 7
- 108010080698 Peptones Proteins 0.000 description 7
- 229940041514 candida albicans extract Drugs 0.000 description 7
- 238000012262 fermentative production Methods 0.000 description 7
- 235000019319 peptone Nutrition 0.000 description 7
- 239000012138 yeast extract Substances 0.000 description 7
- 241000606750 Actinobacillus Species 0.000 description 6
- 235000017557 sodium bicarbonate Nutrition 0.000 description 5
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 5
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 description 4
- 241000186429 Propionibacterium Species 0.000 description 4
- 241000186428 Propionibacterium freudenreichii Species 0.000 description 4
- 239000004744 fabric Substances 0.000 description 4
- 235000010413 sodium alginate Nutrition 0.000 description 4
- 239000000661 sodium alginate Substances 0.000 description 4
- 229940005550 sodium alginate Drugs 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 235000011089 carbon dioxide Nutrition 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 241000948980 Actinobacillus succinogenes Species 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 2
- 229920000728 polyester Polymers 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 210000002268 wool Anatomy 0.000 description 2
- 241000186426 Acidipropionibacterium acidipropionici Species 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000193403 Clostridium Species 0.000 description 1
- 241000272925 Clostridium tyrobutyricum DSM 2637 = ATCC 25755 = JCM 11008 Species 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 241000235527 Rhizopus Species 0.000 description 1
- 240000000111 Saccharum officinarum Species 0.000 description 1
- 235000007201 Saccharum officinarum Nutrition 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000031018 biological processes and functions Effects 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 235000010418 carrageenan Nutrition 0.000 description 1
- 229920001525 carrageenan Polymers 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 238000009418 renovation Methods 0.000 description 1
- 230000000452 restraining effect Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/16—Particles; Beads; Granular material; Encapsulation
- C12M25/18—Fixed or packed bed
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/26—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pH
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/10—Separation or concentration of fermentation products
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Sustainable Development (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Immunology (AREA)
- Analytical Chemistry (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention discloses application of bagasse as an immobilized material in an immobilized fiber bed reactor for producing organic acid by fermentation. The bagasse as an immobilized material can be applied to an internal immobilized fiber bed reactor and an external immobilized fiber bed reactor. In the invention, bagasse used by immobilized cells is waste in sugar industry, and the cost of the bagasse as an immobilized material is extremely low. The specific surface area of the crushed bagasse is increased, cells can be adsorbed on the fiber surface and in fiber gaps in a large amount, and the cells can automatically fall off when the cells are in a decline stage or the thallus activity is poor, so that the cell activity adsorbed on the fibers is ensured. The production efficiency of the organic acid of the immobilized fiber bed reactor using bagasse as an immobilized material is far higher than that of the traditional gel embedding method and also higher than that of other external and built-in plant fiber bed reactors with the highest production efficiency in the current report.
Description
Technical field
The invention belongs to the fermentation engineering field, relate to immobilization fibre bed reactor and produce in the organic acid, bagasse is as the application of immobilization material.
Background technology
Organic acid is meant a class to have carboxyl in molecular structure (compound COOH) is with a wide range of applications in fields such as food, feed, plastics.Traditional organic acid production method mainly is that raw material adopts chemical synthesis to produce with the oil.Current, biological process prepares organic acid because advantages such as product is natural, abundant raw material have been subjected to increasing favor.But still there are a lot of problems in organic acid fermentation production at present.The organic acid microbial fermentation, as ubiquity thalline production in propionic acid, lactic acid, butyric acid and the succsinic acid fermentative production slowly, problem such as lag phase is long.In addition, in the organic acid fermentation production process, the existence of metabolic end product (organic acid) has certain restraining effect to thalline, has limited the further generation of product, all is unfavorable for realizing the organic acid High-efficient Production.
Traditional process for fixation mainly adopts the gel embedding method, is about to cell and is wrapped in the immobilization production that realizes cell in sodium alginate, carrageenin, the agar isogel.Producing propionic acid with immobilization is example, and Woskow SA etc. are by being fixed in propionibacterium in the sodium alginate, and through the 192h fermentation, propionic acid output reaches 47.0g/L, and production efficiency reaches 0.25g/L/h
[1], compare (production efficiency generally is lower than 0.10g/L/h) with the free cell fermentation, have certain advantage.But because the restriction of mass transfer condition, the cell of gel embedding method embedding is difficult to realize self, is difficult to guarantee the thalline vigor for a long time, and is not suitable for the viable cell fermentation.In addition, the cost of gel embedding is higher, is not suitable for organic acid scale operation.
In recent years, be subjected to extensive concern as the cotton fibre bed bioreactor of immobilization material as a kind of novel process for fixation with cotton fibre.Yang ST etc. has made up a kind of external placed type cotton fibre bed bioreactor, and it successfully is used to comprise the multiple organic acid production of propionic acid, lactic acid and butyric acid, has obtained good result.In research process in the past, we have made up a kind of built-in cotton fibre bed bioreactor (Chinese patent publication number: CN101182457A
[3]) and be used for organic acid fermentation, studies show that, this reactor is a kind of effective organic acid production method, with the propionic acid is example, utilize built-in cotton fibre bed bioreactor to produce propionic acid, fermentative production 496h, (Yang ST etc. utilizes external placed type cotton fibre bed bioreactor fermentative production 900h, and propionic acid concentration reaches 71.2g/L can to produce the propionic acid of about 67.1g/L
[2]).In addition, we also produce organic acid with gauze, wool fabric, polyester piece good as immobilization material, have all obtained good experimental result (Chinese patent publication number: CN101182457A
[3]).But, in order further to improve organic acid production efficiency, reduce production costs, still need to seek more natural, immobilization material efficiently.
Bagasse as sugar cane crushing in the sugar industry after remaining waste material, the fiber-enriched structure is a kind of natural immobilization material, but does not see that up to the present it is applied to the report of organic acid fermentation as immobilization material.In addition, bagasse self still contains part sucrose and can be used as carbon source and be applied to organic acid fermentation.
Summary of the invention
Technical problem to be solved by this invention provides bagasse is applied to organic acid fermentation as immobilization material new using value.
For solving the problems of the technologies described above, the technical solution used in the present invention is as follows:
Bagasse in the immobilization fibre bed reactor of fermentation production of organic acid as the application of immobilization material.
Above-mentioned production organic acid immobilization fibre bed reactor both can be that built-in immobilization fibre bed reactor also can be the external placed type immobilization fibre bed reactor.
Above-mentioned organic acid is propionic acid, lactic acid, butyric acid or succsinic acid.
The built-in immobilization fibre bed reactor of fermentation production of organic acid comprises whipping appts, reactor, built-in immobilization post, pH automatic control device and feed supplement device, whipping appts is suspended from the reactor, built-in immobilization post is fixed in reactor bottom and does not influence stirring, bagasse after the pulverizing is filled in the built-in immobilization post as immobilization material, the pH automatic control device links to each other with reactor respectively with the feed supplement device, and reactor top is provided with an inert gas entrance and a tail gas outlet by pipeline.Wherein, described pH automatic control device is by pH probe, pH detector, constant flow pump, alkali lye bottle, air filter and interconnect pipeline and constituted, pH probe is connected on the reactor with fermented liquid in the reactor and contacts, when pH in the reactor be lower than when controlling the pH value, by constant flow pump the alkali lye in the alkali lye bottle is mended automatic control pH in the reactor, air filter is inserted in alkali lye bottle seal place.Wherein, described feed supplement device is made of constant flow pump, feed supplement bottle, air filter and interconnective pipeline, by constant flow pump the substratum in the feed supplement bottle is mended in the reactor by the pipeline that is connected on the reactor, air filter is inserted in feed supplement bottle seal place.
The external placed type immobilization fibre bed reactor of fermentation production of organic acid comprises whipping appts, reactor, external immobilization post, water bath with thermostatic control, pH automatic control device and feed supplement device, whipping appts is suspended from the reactor, external immobilization post links to each other with reactor, the temperature of external immobilization post is controlled by water bath with thermostatic control, to keep in the external immobilization post temperature consistent with reactor temperature, bagasse after the pulverizing is filled in the external immobilization post as immobilization material, the pH automatic control device links to each other with reactor respectively with the feed supplement device, and reactor top is provided with an inert gas entrance and a tail gas outlet by pipeline.Wherein, described pH automatic control device is by pH probe, pH detector, constant flow pump, alkali lye bottle, air filter and interconnect pipeline and constituted, pH probe is connected on the reactor with fermented liquid in the reactor and contacts, when pH in the reactor be lower than when controlling the pH value, by constant flow pump the alkali lye in the alkali lye bottle is mended automatic control pH in the reactor, air filter is inserted in alkali lye bottle seal place.Wherein, described feed supplement device is made of constant flow pump, feed supplement bottle, air filter and interconnective pipeline, by constant flow pump the substratum in the feed supplement bottle is mended in the reactor by the pipeline that is connected on the reactor, air filter is inserted in feed supplement bottle seal place.
In the above-mentioned built-in or external placed type immobilization fibre bed reactor, the alkali lye in the alkali lye bottle can be sodium hydroxide or the 3～8mol/L sodium bicarbonate aqueous solution of 3～8mol/L; Can carbon source (glucose, glycerine etc.) and other nutritive substances (nitrogenous source, inorganic salt etc.) in the feed supplement bottle; The rare gas element that feeds and the tail gas of discharge are: producing the rare gas element that propionic acid, lactic acid or butyric acid feed is nitrogen, and tail gas is the mixture of nitrogen and carbonic acid gas, and the rare gas element that feeds during the production succsinic acid is a carbonic acid gas, and tail gas is carbonic acid gas.
Among the present invention, propionic acid is produced bacterial strain and is propionibacterium commonly used, as: propionibacterium freudenreichii NX-4 (Chinese patent publication number: CN101182457A, culture presevation number: CCTCC No.M207015); Described lactic-acid-producing strain is a lactobacillus, as lactobacillus rhamnosus ATCC 11443; Butyric acid is produced bacterial strain and is butyric acid carboxylic bacterium commonly used, as: junket butyric acid carboxylic bacterium ATCC 25755; The production of succinic acid bacterial strain is succsinic acid actinobacillus commonly used, as: succsinic acid actinobacillus ATCC55618.
Utilize above-mentioned built-in immobilization fibre bed reactor production organic acid method to be: with propionic acid, or lactic acid, or butyric acid, or the production inoculation of succsinic acid is cultivated 12～60h in the reactor that seed culture medium is housed in advance, can realize bacterial strain absorption on the bagasse in built-in immobilization post, seed culture medium in the reactor is changed to fermention medium, utilize the pH automatic control device to mend alkali lye with the fermented liquid pH value in the controlling reactor, starting whipping appts makes the interior material of reactor fully contact the corresponding propionic acid of fermentative production, or lactic acid, or butyric acid, or succsinic acid.
Utilize above-mentioned built-in immobilization fibre bed reactor to produce the organic acid concrete grammar to be: propionic acid or lactic acid or butyric acid or the production of succinic acid bacterial strain inoculum size with 5～20% (v/v) is inoculated in the reactor that the respective seed substratum is housed respectively in advance, feed rare gas element from inert gas entrance, cultivate 12～60h, realize the absorption (being that bacterial strain is fixed on the bagasse) of bacterial strain.Seed liquor in the reactor is replaced with fermention medium, feed rare gas element from inert gas entrance, utilize the pH automatic control device to mend into alkali lye, fermented liquid pH value in the controlling reactor, (the abundant contact of material in mixing speed 0～600r/min) the realization response device realizes the immobilization batch fermentation of propionic acid, lactic acid, butyric acid or production of succinic acid bacterium to utilize whipping appts simultaneously.After cultivating certain hour, when the substrates such as glucose in the fermented liquid consume substantially, utilize constant flow pump to mend the immobilization fed-batch fermentation that to realize propionic acid, lactic acid, butyric acid or succsinic acid into substrates such as glucose from the feed supplement bottle.After cultivating 12～60h, also the seed liquor in the reactor once all can be shifted out, mend the fresh fermention medium of equal volume, realize the immobilization batch fermentation of propionic acid, lactic acid, butyric acid or succsinic acid.
Utilize above-mentioned external placed type immobilization fibre bed reactor production organic acid method to be: with propionic acid, or lactic acid, or butyric acid, or the production of succinic acid inoculation is cultivated 12～60h in the reactor that seed culture medium is housed in advance, by constant flow pump the bacterial strain in the reactor is fully contacted with bagasse in the external immobilization post, bacterial strain is adsorbed on the immobilization that realizes bacterial strain on the bagasse, seed liquor in the reactor is replaced with fermention medium, utilize the pH automatic control device to mend alkali lye with the fermented liquid pH value in the controlling reactor, starting whipping appts makes the interior material of reactor fully contact the corresponding propionic acid of fermentative production, or lactic acid, or butyric acid, or succsinic acid.
Utilize above-mentioned external placed type immobilization fibre bed reactor production organic acid concrete grammar to be: propionic acid, lactic acid, butyric acid or the production of succinic acid bacterial strain inoculum size with 5～20% (v/v) is inoculated in the reactor that the respective seed substratum is housed respectively in advance, feed rare gas element from inert gas entrance, cultivate 12～60h, by constant flow pump the bacterial strain in the reactor is fully contacted with bagasse in the external immobilization post, bacterial strain is adsorbed on the immobilization that realizes bacterial strain on the bagasse.Seed liquor in the reactor is replaced with fermention medium, feed rare gas element from inert gas entrance, utilize the pH automatic control device to mend alkali lye with the fermented liquid pH value in the controlling reactor, utilize the abundant contact of material in whipping appts (mixing speed 0-600r/min) the realization response device simultaneously, realize the immobilization fermentation of propionic acid, lactic acid, butyric acid or production of succinic acid bacterium.After cultivating certain hour, when the substrates such as glucose in the fermented liquid consume substantially, utilize constant flow pump to mend the immobilization fed-batch fermentation that to realize propionic acid, lactic acid, butyric acid or succsinic acid into substrates such as glucose from the feed supplement bottle.After cultivating 12～60h, also the seed liquor in the reactor once all can be shifted out, mend the fresh fermention medium of equal volume, realize the immobilization batch fermentation of propionic acid, lactic acid, butyric acid or succsinic acid.
Compare with other immobilization materials, utilize bagasse to have following advantage as immobilization material:
1, the cell fixation cost is extremely low: the used bagasse of immobilized cell is the waste material in the sugar industry, and it is as immobilization material cost extremely low (200～500 yuan/ton of market prices).General gel embedding method embedded material is expensive (as sodium alginate, 26000～35000 yuan/ton of market prices) comparatively, also is that price is high and compare with bagasse through artificial fabric with cotton, gauze, wool fabric, polyester piece good etc.
2, the efficient absorption and the continual renovation of cell: bagasse specific surface area after crushed increases, cell can be adsorbed in fiber surface and the fibre gap in a large number, when cell is in the paracme or the thalline vigor is relatively poor, be difficult to be attached to fiber surface, cell will show as automatically and come off, the adsorption site of vacating can be adsorbed on the bagasse for the new fresh cell of high vigor again, the decline of bacterial strain vigor between fermentation strain can too much not cause batch because batch fermentation, thus the stability of batch fermentation organic acid production efficiency guaranteed.
In the fermenting process, thalline vigor and organic acid fermentation are closely related.Traditional gel embedding method cell is embedded in and is used for organic acid fermentation in the gel, and thalline is along with the nature that increases of batch fermentation fails, and vigor reduces.But because the constraint of gel, the cell that is in the paracme still is stranded in the intercellular substance, can not free in and out, and vigor is difficult to guarantee.And with the fabric that comprises cotton fibre during as immobilization material cell can freely come off and upgrade, but compare with bagasse, the former specific surface area is less, is adsorbed on the fiber cell total amount than few slightly in the bagasse.
3, organic acid better tolerance, production efficiency height: organic acid has the intensive product inhibition for the fermentation of thalline, and output will be difficult to further raising when organic acid reaches finite concentration, has greatly hindered the organic acid fermentation.And with bagasse be the organic acid production efficiency of immobilization fibre bed reactor of immobilization material far above traditional gel embedding method, also be higher than production efficiency is the highest in the present report external placed type and built-in cotton fibre bed bioreactor (table 1～table 4).With the fermentation productions of ethylformic acid is example, the propionic acid concentration that traditional gel embedding method is produced is difficult to break through 50g/L and (utilizes this method to produce the highest 47.0g/L of being reported as of propionic acid concentration at present, production efficiency is 0.25g/L/h, see document 1), (this method is produced the highest 71.2g/L of being reported as of propionic acid at present and be difficult to break through 75g/L with cotton fibre reactor propionic fermentation concentration, production efficiency is 0.08g/L/h, see document 2), and utilize bagasse as immobilization material, in this research process, utilizing external placed type and built-in vegetable fibre bed bioreactor propionic acid concentration to distinguish can be up to 132.5g/L, 124.1g/L, production efficiency is respectively 0.66g/L/h, 0.62g/L/h visible two kinds of production methods are propionic acid production method efficiently.
Table 1. different fixing method is produced propionic acid relatively
| Process for fixation | Immobilization material | Propionic acid concentration (g/L) | Fermentation time (h) | Production efficiency (g/L/h) | Reference |
| The gel embedding method | Sodium alginate | 47.0 | 192 | 0.25 | [1] |
| External placed type cotton fibre bed bioreactor | Cotton fibre | 71.2 | 900 | 0.08 | [2] |
| Built-in cotton fibre bed bioreactor | Cotton fibre | 67.1 | 496 | 0.14 | [3] |
| External placed type vegetable fibre bed bioreactor | Bagasse | 132.5 | 200 | 0.66 | The present invention |
| Built-in vegetable fibre bed bioreactor | Bagasse | 124.1 | 200 | 0.62 | The present invention |
Table 2. different fixing method is produced L-lactic acid relatively
| Process for fixation | Immobilization material | Lactic acid concn (g/L) | Fermentation time (h) | Production efficiency (g/L/h) | Reference |
| Rotary cotton fibre bed bioreactor | Cotton fibre | 127.0 | 54 | 2.35 | [4] |
| External placed type vegetable fibre bed bioreactor | Bagasse | 198.9 | 80 * | 2.49 | The present invention |
| Built-in vegetable fibre bed bioreactor | Bagasse | 182.2 | 80 * | 2.27 | The present invention |
Annotate
*: lactic acid concn is bigger for the fermentation time influence, utilizes external placed type and built-in vegetable fibre bed bioreactor to produce L-lactic acid, and the L-lactic acid concn can reach 127.0g/L behind ferment respectively 40h, the 43h, and this moment, production efficiency was respectively 3.18g/L/h and 2.95g/L/h.
Table 3. different fixing method is produced butyric acid relatively
| Process for fixation | Immobilization material | Butyric acid density (g/L) | Fermentation time (h) | Production efficiency (g/L/h) | Reference |
| External placed type cotton fibre bed bioreactor | Cotton fibre | 49.9 | 260 | 0.19 | [5] |
| Built-in cotton fibre bed bioreactor | Cotton fibre | 35.5 | 80 | 0.44 | [3] |
| External placed type vegetable fibre bed bioreactor | Bagasse | 75.5 | 80 | 0.94 | The present invention |
| Built-in vegetable fibre bed bioreactor | Bagasse | 71.6 | 80 | 0.89 | The present invention |
Table 4. different fixing method is produced succsinic acid relatively
| Process for fixation | Immobilization material | Succsinic acid concentration (g/L) | Fermentation time (h) | Production efficiency (g/L/h) | Reference |
| Built-in cotton fibre bed bioreactor | Cotton fibre | 100.0 | 48 | 2.08 | [3] |
| External placed type vegetable fibre bed bioreactor | Bagasse | 120.0 | 52 | 2.31 | The present invention |
| Built-in vegetable fibre bed bioreactor | Bagasse | 112.3 | 52 | 2.16 | The present invention |
Reference:
[1]Woskow?SA.，Glatz?BA.Propionic?acid?production?by?a?propionic?acid-tolerant?strain?ofPropionibacterium?acidipropionici?in?batch?and?semicontinuous?fermentation.ApplEnviron?Microbiol1991；57(10)：2821-2828.
[2]Suwannakham?S，Yang?ST. Enhanced?propionic?acid?fermentation?by?Propionibacteriumacidipropionici?mutant?obtained?by?adap
[3] Xu Hong, Feng Xiaohai, Li Kai, Li Sha, Ouyang Pingkai. the immobilization fibre bed reactor of a kind of fermentation productions of ethylformic acid, butyric acid, succsinic acid [P]. publication number: CN101182457A.2008.5.21.
[4]Tay?A，Yang?ST.Production?of?L(+)-lactic?acid?from?glucose?and?starch?by?immobilizedcells?of?Rhizopus?oryzae?in?a?rotating?fibrous?bed?bioreactor.Biotechnol?Bioeng?2002；80(1)：1-12.
[5]Xiaoguang?Liu，Shang-Tian?Yang.Kinetics?of?butyric?acid?fermentation?of?glucose?andxylose?by?Clostridium?tyrobutyricum?wild?type?and?mutant[J].Process?Biochem?2006，41：801-808.
Description of drawings
Fig. 1 is the structural representation of built-in immobilization fibre bed reactor of the present invention.
Wherein, 1-1. air filter, 1-2. air filter, 1-3. air filter, 2. alkali lye bottle, 3. feed supplement bottle, 4. inert gas entrance, 5-1. constant flow pump, 5-2. constant flow pump, 6.pH detector, 7.pH probe, 8. temp probe, 9. whipping appts, 10. reactor, the built-in immobilization post of 11a. (including bagasse), the outlet of 12. tail gas.
Fig. 2 is the structural representation of external placed type immobilization fibre bed reactor of the present invention.
Wherein, 1-1. air filter, 1-2. air filter, 1-3. air filter, 2. alkali lye bottle, 3. feed supplement bottle, 4. inert gas entrance, 5-1. constant flow pump, 5-2. constant flow pump, 5-3. constant flow pump, 6.pH detector, 7.pH probe, 8. temp probe, 9. whipping appts, 10. reactor, 11b. external immobilization post (including bagasse), the outlet of 12. tail gas, 13. waters bath with thermostatic control.
Embodiment:
According to following embodiment, the present invention may be better understood.Yet, those skilled in the art will readily understand that the described concrete material proportion of embodiment, processing condition and result thereof only are used to illustrate the present invention, and should also can not limit the present invention described in detail in claims.
Fig. 1 as seen, built-in immobilization fibre bed reactor of the present invention comprises whipping appts 9, reactor 10, built-in immobilization post 11a, pH automatic control device and feed supplement device; Whipping appts 9 is suspended from the reactor 10; Built-in immobilization post 11a is fixed in reactor 10 bottoms and does not influence stirring, built-in immobilization post 11a is cylindric, the surface is covered with aperture (about the about 1mm in aperture), and diameter is about 1/5 of reactor diameter, and the bagasse after the pulverizing is filled among the built-in immobilization post 11a as immobilization material; The pH automatic control device links to each other with reactor 10 respectively with the feed supplement device; Temp probe 8 places reactor 10, contacts with fermented liquid.Wherein, described pH automatic control device is by pH probe 7, pH detector 6, constant flow pump 5-1, alkali lye bottle 2, air filter 1-1 and interconnect pipeline and constituted, pH probe 7 is connected on the reactor 10 and contacts with fermented liquid, when pH in the reactor 10 be lower than when controlling the pH value, by constant flow pump 5-1 the alkali lye in the alkali lye bottle 2 is mended automatic control pH in the reactor 10, air filter 1-1 is inserted in alkali lye bottle 2 seal places.Wherein, described feed supplement device is made of constant flow pump 5-2, feed supplement bottle 3, air filter 1-2 and interconnective pipeline, by constant flow pump 5-2 the substratum in the feed supplement bottle 3 is mended in the reactor 10 by the pipeline that is connected on the reactor 10, air filter 1-2 is inserted in feed supplement bottle 3 seal places.The built-in immobilization fibre bed reactor that uses in following examples is all reactor shown in Figure 1.
As seen from Figure 2, external placed type immobilization fibre bed reactor of the present invention comprises whipping appts 9, reactor 10, external immobilization post 11b, water bath with thermostatic control 13, pH automatic control device and feed supplement device; Whipping appts 9 is suspended from the reactor 10; External immobilization post 11b links to each other with reactor 10, the temperature of external immobilization post 11b is by water bath with thermostatic control 13 controls, to keep in the external immobilization post 11b in the temperature and reactor 10 temperature consistent, bagasse after the pulverizing is filled in the external immobilization post 11b as immobilization material, external immobilization post 11b is sealed by stainless (steel) wire at two ends, and bagasse can not spill; The pH automatic control device links to each other with reactor 10 respectively with the feed supplement device; Temp probe 8 places reactor 10, contacts with fermented liquid.Wherein, described pH automatic control device is by pH probe 7, pH detector 6, constant flow pump 5-1, alkali lye bottle 2, air filter 1-1 and interconnect pipeline and constituted, pH probe 7 is connected on the reactor 10 and contacts with fermented liquid, when pH in the reactor 10 be lower than when controlling the pH value, by constant flow pump 5-1 the alkali lye in the alkali lye bottle 2 is mended automatic control pH in the reactor 10, air filter 1-1 is inserted in alkali lye bottle 2 seal places.Wherein, described feed supplement device is made of constant flow pump 5-2, feed supplement bottle 3, air filter 1-2 and interconnective pipeline, by constant flow pump 5-2 the substratum in the feed supplement bottle 3 is mended in the reactor 10 by the pipeline that is connected on the reactor 10, air filter 1-2 is inserted in feed supplement bottle 3 seal places.The external placed type immobilization fibre bed reactor that uses in following examples is all reactor shown in Figure 2.
Utilize propionibacterium freudenreichii NX-4 (the Propionibacterium freudeenreichiiNX-4 of this laboratory patent applied for, Chinese patent publication number: CN101182457A, culture presevation number: CCTCC No.M207015) in built-in vegetable fibre bed bioreactor, produce propionic acid.Seed culture medium: glucose 20g/L, peptone 5g/L, yeast extract paste 5g/L, NaCl5g/L, pH6.9.Fermention medium: peptone 10g/L, yeast extract paste 5g/L, NaCl 3g/L, (NH
4)
2SO
45g/L, KH
2PO
45g/L, pH6.9.
Bagasse is fixed among the built-in immobilization post 11a, propionibacterium freudenreichii NX-4 seed liquor is inserted (cumulative volume: 4L) in the reactor 10 of the seed culture medium that the 3.6L volume is housed in advance by the inoculum size (0.4L) of 10% (v/v), 35 ℃ of temperature feed 0.1L/min N
2Utilize whipping appts 9 to stir (rotating speed 100r/min), promote the continuous exchange of the inside and outside liquid of built-in immobilization post 11a, utilize constant flow pump 5-1 to mend pH6.0 in the 8mol/L sodium hydroxide solution controlling reactor 10 simultaneously, cultivate 36h and realize the immobilization of cell on bagasse fibre.Seed liquor in the reactor 10 is replaced by fermention medium (initial glucose concentration 40g/L is that glucose is added in the fermention medium), initial pH is 6.9, utilize constant flow pump 5-1 to mend pH6.0 in the 8mol/L sodium hydroxide solution controlling reactor 10,35 ℃ of temperature feed 0.1L/min N
2, utilize whipping appts 9 to stir (100r/min) simultaneously.When being lower than 10g/L, glucose concn the glucose in the feed supplement bottle 3 (500g/L) is mended in the reactor 10 by constant flow pump 5-2, it is 35-40g/L that single is mended into the back glucose concn, feeding glucose fermentation 200h (glucose total concn: 200g/L) successively, propionic acid output 93.8g/L, production efficiency reaches 0.47g/L/h.
Bacterial classification, culture condition and feed supplement mode are with embodiment 1, and different is that immobilization material changes cotton fibre into by bagasse.Utilize built-in cotton fibre bed bioreactor to produce propionic acid, behind the fermentative production 496h, propionic acid output 67.0g/L, production efficiency is 0.14g/L/h.
Embodiment 3 built-in vegetable fibre bed bioreactors are produced propionic acid
Bacterial classification, culture condition are with embodiment 1.Initial glucose concentration is 100g/L, when glucose concn is lower than 10g/L, by constant flow pump 5-2 mend once more into 80g/L glucose (the glucose total concn: 180g/L), the fermentation 350h, propionic acid output 74.3g/L, production efficiency reaches 0.21g/L/h.
Bacterial classification, culture condition are with embodiment 1.Initial glucose concentration is 40g/L, when being lower than 10g/L, the glucose in the feed supplement bottle 3 (500g/L) constant current is mended in the reactor 10 glucose concn by constant flow pump 5-2, concentration in the controlling reactor 10 is 10g/L, fermentation 200h (glucose total concn: 260g/L), propionic acid output 124.1g/L, production efficiency reaches 0.62g/L/h.
Embodiment 5 built-in vegetable fibre bed bioreactors are produced propionic acid
Bacterial classification, culture condition and feed supplement mode are with embodiment 1.Single batch of glucose total concn is controlled to be 200g/L, and fermentation is replaced fresh culture behind the 200h, ferment 20 batches altogether behind the 4000h unit volume (L) produce propionic acid meter 1845.0g altogether, single batch of propionic acid concentration reaches 92.3g/L, production efficiency reaches 0.46g/L/h.
Embodiment 6 external placed type vegetable fibre bed bioreactors are produced propionic acid
Bacterial classification, culture condition and feed supplement mode are with embodiment 1.Bagasse is fixed among the external immobilization post 11b, by the inoculum size (0.4L) of 10% (v/v) propionibacterium freudenreichii NX-4 seed liquor is inserted and be equipped with in advance in the reactor 10 of 3.6L seed culture medium, 35 ℃ of temperature feed 0.1L/minN
2Utilize whipping appts 9 to stir (rotating speed 100r/min), utilize constant flow pump 5-1 to mend pH6.0 in the 8mol/L sodium hydroxide solution controlling reactor 10 simultaneously, utilize the continuous exchange of liquid in constant flow pump realization response device 10 and the immobilization post 11b, the temperature of immobilization post 11b is controlled to be 35 ℃ through water bath with thermostatic control 13, cultivates 36h and realizes the immobilization of cell on bagasse.Seed liquor in the reactor 10 is replaced by fermention medium (initial glucose concentration 40g/L is that glucose is added in the fermention medium), initial pH is 6.9, utilize constant flow pump 5-1 to mend pH6.0 in the 8mol/L sodium hydroxide solution controlling reactor 10,35 ℃ of temperature feed 0.1L/min N
2, utilize whipping appts 9 to stir (100r/min) simultaneously.When being lower than 10g/L, glucose concn the glucose in the feed supplement bottle 3 (500g/L) is mended in the reactor 10 by constant flow pump 5-2, it is 35-40g/L that single is mended into the back glucose concn, feeding glucose fermentation 200h (glucose total concn: 260g/L) successively, propionic acid output 119.2g/L, production efficiency reaches 0.60g/L/h.
Embodiment 7 external placed type cotton fibre bed bioreactors are produced propionic acid (reference examples 2)
Bacterial classification, culture condition and feed supplement mode are with embodiment 1, and different is that immobilization material changes cotton fibre into by bagasse.Utilize external placed type cotton fibre bed bioreactor to produce propionic acid, behind the fermentative production 496h, propionic acid output 59.18g/L, people such as this test-results and Yang ST utilize external placed type cotton fibre bed bioreactor to produce propionic acid concentration basically identical (bacterial strain uses therefor: acid propionibacterium ATCC 4875, contrast time: 496h).
Bacterial classification, culture condition and feed supplement mode are with embodiment 6.Single batch of glucose total concn is controlled to be 200g/L, and fermentation is replaced fresh culture behind the 140h, ferment 20 batches altogether behind the 2800h unit volume (L) produce propionic acid meter 1950.0g altogether, single batch of propionic acid concentration reaches 97.5g/L, production efficiency reaches 0.70g/L/h.
Bacterial classification, culture condition are with embodiment 6.Initial glucose concentration is 100g/L, when glucose concn is lower than 10g/L, by constant flow pump 5-2 mend once more into 80g/L glucose (the glucose total concn: 180g/L), the fermentation 300h, propionic acid output 78.2g/L, production efficiency reaches 0.26g/L/h.
Bacterial classification, culture condition are with embodiment 6.Initial glucose concentration is 40g/L, when being lower than 10g/L, the glucose in the feed supplement bottle 3 (500g/L) constant current is mended in the reactor 10 glucose concn by constant flow pump 5-2, concentration in the controlling reactor 10 is 10g/L, fermentation 200h (glucose total concn: 275g/L), propionic acid output 132.5g/L, production efficiency reaches 0.66g/L/h.
Embodiment 11 built-in vegetable fibre bed bioreactors are produced L-lactic acid
Utilize lactobacillus rhamnosus ATCC11443 (Lactobacillus rhamnosus ATCC11443) in built-in vegetable fibre bed bioreactor, to produce L-lactic acid.Seed culture medium: glucose 50g/L, yeast extract paste 10g/L, peptone 5g/L, sodium acetate 0.5g/L, MgSO
40.15g/L, MnSO
40.01g/L, FeSO
40.01g/L, pH6.0.Fermention medium: corn steep liquor 25g/L, sodium acetate 5g/L, MgSO
40.15g/L, MnSO
40.01g/L, FeSO
40.01g/L, tween 80 1mL, pH6.0.Bagasse is fixed among the built-in immobilization post 11a, by the inoculum size (0.16L) of 4% (v/v) lactobacillus rhamnosus ATCC 11443 seed liquor is inserted and be equipped with in advance in the reactor 10 of 3.84L seed culture medium, 37 ℃ of temperature feed 0.2L/min N
2, utilize whipping appts 9 to stir (rotating speed 200r/min) simultaneously, utilize constant flow pump 5-1 to mend 6mol/L sodium hydroxide solution control pH6.0 and cultivate the immobilization of 24h realization cell on bagasse.Seed liquor in the reactor 10 is replaced by fermention medium (initial glucose 40g/L), and 37 ℃ of temperature utilize constant flow pump 5-1 to mend into 6mol/L sodium hydroxide solution control pH6.0, feed 0.2L/min N
2, utilize whipping appts 9 to stir (rotating speed 200r/min) fermentation simultaneously.When being lower than 10g/L, glucose concn the glucose in the feed supplement bottle 3 (500g/L) is mended in the reactor 10 by constant flow pump 5-2, control mends the back glucose concn at 35-40g/L, feeding glucose fermentation 80h successively, L-lactic acid production 182.2g/L (glucose total concn 200g/L), production efficiency reaches 2.27g/L/h.
Bacterial classification, culture condition are with embodiment 11.Utilize lactobacillus rhamnosus ATCC 11443 (Lactobacillusrhamnosus ATCC 11443) in external placed type vegetable fibre bed bioreactor, to produce L-lactic acid.Bagasse is fixed among the external immobilization post 11b, by the inoculum size (0.2L) of 5% (v/v) lactobacillus rhamnosus ATCC 11443 seed liquor is inserted and be equipped with in advance in the reactor 10 of 3.8L seed culture medium, 37 ℃ of temperature feed 0.2L/min N
2, utilize whipping appts 9 to stir (rotating speed 200r/min) simultaneously, utilize constant flow pump 5-1 to mend 6mol/L sodium hydroxide solution control pH6.0 and cultivate the immobilization of 24h realization cell on bagasse.Seed liquor in the reactor 10 is replaced by fermention medium (initial glucose 40g/L), and 37 ℃ of temperature utilize constant flow pump 5-1 to mend into 6mol/L sodium hydroxide solution control pH6.0, feed 0.2L/min N
2, utilize whipping appts 9 to stir (rotating speed 200r/min) fermentation simultaneously.When being lower than 10g/L, glucose concn the glucose in the feed supplement bottle 3 (500g/L) is mended in the reactor 10 by constant flow pump 5-2, control mends the back glucose concn at 35-40g/L, feeding glucose fermentation 80h successively, L-lactic acid production 198.9g/L (glucose total concn 220g/L), production efficiency reaches 2.49g/L/h.
Utilize junket butyric acid carboxylic bacterium ATCC 25755 (Clostridium tyrobutyricum ATCC 25755) in built-in vegetable fibre bed bioreactor, to produce butyric acid.Seed culture medium: glucose 20g/L, yeast extract paste 3g/L, peptone 1g/L, (NH
4)
2SO
41g/L, K
2HPO
41g/L, pH6.0.Fermention medium: yeast extract paste 5g/L, peptone 3g/L, (NH
4)
2SO
41.5g/L, K
2HPO
41g/L, MgSO
40.1g/L, pH6.0.Bagasse is fixed among the built-in immobilization post 11a, by the inoculum size (0.2L) of 5% (v/v) junket butyric acid carboxylic bacterium ATCC 25755 seed liquor is inserted and be equipped with in advance in the reactor 10 of 3.8L seed culture medium, 37 ℃ of temperature feed 0.2L/min N
2, utilize whipping appts 9 to stir (rotating speed 150r/min) simultaneously, utilize constant flow pump 5-1 to mend 6mol/L sodium hydroxide solution control pH6.3 and cultivate the immobilization of 24h realization cell on bagasse.Seed liquor in the reactor 10 is replaced by fermention medium (initial glucose 40g/L), and 37 ℃ of temperature utilize constant flow pump 5-1 to mend into 6mol/L sodium hydroxide solution control pH6.3, feed 0.2L/min N
2, utilize whipping appts 9 to stir (rotating speed 150r/min) fermentation simultaneously.When being lower than 10g/L, glucose concn the glucose in the feed supplement bottle 3 (500g/L) is mended in the reactor 10 by constant flow pump 5-2, control mends the back glucose concn at 35-40g/L, feeding glucose fermentation 80h successively, butyric acid output 71.6g/L (glucose total concn 170g/L), production efficiency reaches 0.89g/L/h.
Embodiment 14 external placed type vegetable fibre bed bioreactors are produced butyric acid
Bacterial classification, culture condition are with embodiment 13.Bagasse is fixed among the external immobilization post 11b, junket butyric acid carboxylic bacterium ATCC 25755 (0.32L) seed liquor is inserted in the reactor 10 of the seed culture medium that 3.68L is housed in advance by the inoculum size of 8% (v/v), 37 ℃ of temperature feed 0.2L/minN
2, utilize whipping appts 9 to stir (rotating speed 150r/min) simultaneously, utilize constant flow pump 5-1 to mend 6mol/L sodium hydroxide solution control pH6.3 and cultivate the immobilization of 24h realization cell on bagasse.Seed liquor in the reactor 10 is replaced by fermention medium (initial glucose 40g/L), and 37 ℃ of temperature utilize constant flow pump 5-1 to mend into 6mol/L sodium hydroxide solution control pH6.3, feed 0.2L/min N
2, utilize whipping appts 9 to stir (rotating speed 150r/min) fermentation simultaneously.When being lower than 10g/L, glucose concn the glucose in the feed supplement bottle 3 (500g/L) is mended in the reactor 10 by constant flow pump 5-2, control mends the back glucose concn at 35-40g/L, feeding glucose fermentation 80h successively, butyric acid output 75.5g/L (glucose total concn 180g/L), production efficiency reaches 0.94g/L/h.
Embodiment 15 built-in vegetable fibre bed bioreactors are produced succsinic acid
Utilize succsinic acid actinobacillus ATCC 55618 (Actinobacillus succinogenes ATCC 55618) in immobilization fibre bed reactor, to produce succsinic acid.Seed culture medium: glucose 20g/L, yeast extract paste 5g/L, peptone 1g/L, (NH
4)
2SO
41g/L, K
2HPO
41g/L, NaCl0.5g/L, pH6.5.Fermention medium: yeast extract paste 10g/L, peptone 5g/L, Na
2HPO
41g/L, Na
2KHPO
41g/L, MgCl
20.2g/L, pH6.5.Bagasse is fixed among the built-in immobilization post 11a, succsinic acid actinobacillus ATCC 55618 seed liquor is inserted in the reactor 10 of the seed culture medium that 3.6L is housed in advance by the inoculum size (0.4L) of 10% (v/v), 37 ℃ of temperature feed 0.5L/min CO
2, utilize whipping appts 9 to stir (rotating speed 100r/min), utilize constant flow pump 5-1 to mend simultaneously into 6mol/L sodium hydrogen carbonate solution control pH6.0, cultivate 24h and realize the immobilization of cell on fiber.Seed liquor in the fibre bed reactor is replaced by fermention medium, initial glucose 100g/L, 37 ℃ of temperature utilize constant flow pump 5-1 to mend into 6mol/L sodium hydrogen carbonate solution control pH6.0, feed 1.0L/min CO
2Utilize whipping appts 9 to stir (rotating speed 100r/min) simultaneously, when being lower than 10g/L, glucose concn the glucose in the feed supplement bottle 3 (500g/L) is mended in the reactor 10 by constant flow pump 5-2, control mends the back glucose concn at 35-40g/L, feeding glucose fermentation 52h (glucose total concn 160g/L) successively, succinic acid production 112.3g/L, production efficiency reaches 2.16g/L/h.
Embodiment 16 external placed type vegetable fibre bed bioreactors are produced succsinic acid
Bacterial classification, culture condition are with embodiment 15.Utilize succsinic acid actinobacillus ATCC 55618 (Actinobacillussuccinogenes ATCC 55618) in immobilization fibre bed reactor, to produce succsinic acid.Bagasse is fixed among the external immobilization post 11b, by the inoculum size (0.4L) of 10% (v/v) succsinic acid actinobacillus ATCC 55618 seed liquor is inserted and be equipped with in advance in the reactor 10 of 3.6L seed culture medium, 37 ℃ of temperature feed 0.5L/min CO
2, utilize whipping appts 9 to stir (rotating speed 100r/min), utilize constant flow pump 5-1 to mend 6mol/L sodium hydrogen carbonate solution control pH6.0 simultaneously and cultivate the immobilization of 24h realization cell on bagasse.Seed liquor in the fibre bed reactor is replaced by fermention medium, initial glucose 100g/L, 37 ℃ of temperature utilize constant flow pump 5-1 to mend into 6mol/L sodium hydrogen carbonate solution control pH6.0, feed 1.0L/min CO
2Utilize whipping appts 9 to stir (rotating speed 100r/min) simultaneously, when being lower than 10g/L, glucose concn the glucose in the feed supplement bottle 3 (500g/L) is mended in the reactor 10 by constant flow pump 5-2, control mends the back glucose concn at 35-40g/L, feeding glucose fermentation 52h (glucose total concn 170g/L) successively, succinic acid production 120.0g/L, production efficiency reaches 2.31g/L/h.
Claims (10)
- Bagasse in the immobilization fibre bed reactor of fermentation production of organic acid as the application of immobilization material.
- 2. application according to claim 1 is characterized in that bagasse application as immobilization material in the built-in immobilization fibre bed reactor of fermentation production of organic acid.
- 3. application according to claim 2, the built-in immobilization fibre bed reactor that it is characterized in that described fermentation production of organic acid comprises whipping appts (9), reactor (10), built-in immobilization post (11a), pH automatic control device and feed supplement device, whipping appts (9) is suspended from the reactor (10), built-in immobilization post (11a) is fixed in reactor (10) bottom and does not influence stirring, bagasse after the pulverizing is filled in the built-in immobilization post (11a) as immobilization material, and the pH automatic control device links to each other with reactor (10) respectively with the feed supplement device.
- 4. application according to claim 3, it is characterized in that described pH automatic control device is by pH probe (7), pH detector (6), constant flow pump (5-1), alkali lye bottle (2), air filter (1-1) and interconnect pipeline and constituted, pH probe (7) is connected to reactor (10) fermented liquid last and in the reactor (10) and contacts, when pH in the reactor (10) be lower than when controlling the pH value, by constant flow pump (5-1) alkali lye in the alkali lye bottle (2) is mended automatic control pH in the reactor (10), air filter (1-1) is inserted in alkali lye bottle (2) seal place.
- 5. application according to claim 3, it is characterized in that described feed supplement device is made of constant flow pump (5-2), feed supplement bottle (3), air filter (1-2) and interconnective pipeline, by constant flow pump (5-2) substratum in the feed supplement bottle (3) is mended in the reactor (10) by the pipeline that is connected on the reactor (10), air filter (1-2) is inserted in feed supplement bottle (3) seal place.
- 6. application according to claim 1 is characterized in that bagasse application as immobilization material in the external placed type immobilization fibre bed reactor of fermentation production of organic acid.
- 7. application according to claim 6, the external placed type immobilization fibre bed reactor that it is characterized in that described fermentation production of organic acid comprises whipping appts (9), reactor (10), external immobilization post (11b), water bath with thermostatic control (13), pH automatic control device and feed supplement device, whipping appts (9) is suspended from the reactor (10), external immobilization post (11b) links to each other with reactor (10), the temperature of external immobilization post (11b) is controlled by water bath with thermostatic control (13), to keep the interior temperature of external immobilization post (11b) consistent with the interior temperature of reactor (10), bagasse after the pulverizing is filled in the external immobilization post (11b) as immobilization material, and the pH automatic control device links to each other with reactor (10) respectively with the feed supplement device.
- 8. application according to claim 7, it is characterized in that described pH automatic control device is by pH probe (7), pH detector (6), constant flow pump (5-1), alkali lye bottle (2), air filter (1-1) and interconnect pipeline and constituted, pH probe (7) is connected to reactor (10) fermented liquid last and in the reactor (10) and contacts, when pH in the reactor (10) be lower than when controlling the pH value, by constant flow pump (5-1) alkali lye in the alkali lye bottle (2) is mended automatic control pH in the reactor (10), air filter (1-1) is inserted in alkali lye bottle (2) seal place.
- 9. application according to claim 7, it is characterized in that described feed supplement device is made of constant flow pump (5-2), feed supplement bottle (3), air filter (1-2) and interconnective pipeline, by constant flow pump (5-2) substratum in the feed supplement bottle (3) is mended in the reactor (10) by the pipeline that is connected on the reactor (10), air filter (1-2) is inserted in feed supplement bottle (3) seal place.
- 10. according to any described application in the claim 1～9, it is characterized in that described organic acid is propionic acid, lactic acid, butyric acid or succsinic acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810234514.8A CN101402914B (en) | 2008-11-20 | 2008-11-20 | Application of bagasse as immobilized material in immobilized fiber bed reactor for producing organic acid by fermentation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810234514.8A CN101402914B (en) | 2008-11-20 | 2008-11-20 | Application of bagasse as immobilized material in immobilized fiber bed reactor for producing organic acid by fermentation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101402914A CN101402914A (en) | 2009-04-08 |
| CN101402914B true CN101402914B (en) | 2011-08-31 |
Family
ID=40537079
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200810234514.8A Expired - Fee Related CN101402914B (en) | 2008-11-20 | 2008-11-20 | Application of bagasse as immobilized material in immobilized fiber bed reactor for producing organic acid by fermentation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101402914B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103146571A (en) * | 2013-04-01 | 2013-06-12 | 南京工业大学 | Column type immobilization reactor for producing rhamnose gum through fermentation and process thereof |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100939236B1 (en) * | 2009-06-22 | 2010-01-29 | 유희룡 | Bagasse composite, composite manufacturing method and interior composite using the same |
| CN102140445B (en) * | 2010-12-31 | 2014-09-03 | 浙江海洋学院 | Carrier cell immobilization method |
| CN103194374B (en) * | 2013-04-01 | 2014-12-31 | 南京工业大学 | Column type immobilization reactor for producing gamma-polyglutamic acid by fermentation and process thereof |
| CN103355728A (en) * | 2013-07-01 | 2013-10-23 | 福建省农业科学院农业工程技术研究所 | Immobilized semi-continuous lactic acid fermentation method of juice |
| CN103555777B (en) * | 2013-11-15 | 2015-07-01 | 厦门大学 | Method for producing lactic acid by using immobilized cells for converting glycerin |
| CN104178421B (en) * | 2014-08-25 | 2016-01-13 | 鲁东大学 | A kind of is the method for fluidized bed type enzyme reactor by stirred-tank fermenter system assembles |
| CN105695370B (en) * | 2016-04-13 | 2019-06-11 | 南京工业大学 | Clostridium butyricum and culture method and application thereof |
| CN106191141A (en) * | 2016-06-13 | 2016-12-07 | 杭州贝谷生物科技有限公司 | The method utilizing feedstuff fibre bed bioreactor fermenting and producing butanoic acid |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101182456A (en) * | 2007-11-29 | 2008-05-21 | 南京工业大学 | Immobilized fibrous bed reactor for producing propionic acid, butyric acid and succinic acid by fermentation |
-
2008
- 2008-11-20 CN CN200810234514.8A patent/CN101402914B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101182456A (en) * | 2007-11-29 | 2008-05-21 | 南京工业大学 | Immobilized fibrous bed reactor for producing propionic acid, butyric acid and succinic acid by fermentation |
Non-Patent Citations (2)
| Title |
|---|
| Jianliang Yu et al.An novel immobilization method of Saccharomyces cerevisiae to sorghum bagasse for ethanol production.《Journal of Biotechnology》.2007,第129卷(第3期),415-420. * |
| Suwannakham S et al.Enhanced propionic acid fermentation by Propionibacterium acidipropionici mutant obtained by adaptation in a fibrous-bed bioreactor.《Biotechnol Bioeng》.2005,第91卷(第3期),325-337. * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103146571A (en) * | 2013-04-01 | 2013-06-12 | 南京工业大学 | Column type immobilization reactor for producing rhamnose gum through fermentation and process thereof |
| CN103146571B (en) * | 2013-04-01 | 2014-08-27 | 南京工业大学 | Column type immobilization reactor for producing rhamnose gum through fermentation and process thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101402914A (en) | 2009-04-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101402914B (en) | Application of bagasse as immobilized material in immobilized fiber bed reactor for producing organic acid by fermentation | |
| Soccol et al. | Potential of solid state fermentation for production of L (+)-lactic acid by Rhizopus oryzae | |
| Xiaodong et al. | Direct fermentative production of lactic acid on cassava and other starch substrates | |
| Feng et al. | Biohydrogen production from apple pomace by anaerobic fermentation with river sludge | |
| John et al. | Simultaneous saccharification and fermentation of cassava bagasse for L-(+)-lactic acid production using Lactobacilli | |
| Roble et al. | L-Lactic acid production from raw cassava starch in a circulating loop bioreactor with cells immobilized in loofa (Luffa cylindrica) | |
| Liang et al. | Enhanced propionic acid production from Jerusalem artichoke hydrolysate by immobilized Propionibacterium acidipropionici in a fibrous-bed bioreactor | |
| CN101182456B (en) | Immobilized fibrous bed reactor for producing propionic acid, butyric acid and succinic acid by fermentation | |
| CN101215584B (en) | Technique for preparing succinic acid by biological transformation of agronomic crop straw | |
| CN102174602B (en) | Method for producing L-lactic acid through biomass fermentation | |
| CN101285047B (en) | High-optical-purity D-lactic acid producing strain and process for producing D-lactic acid by fermentation of same | |
| Zhao et al. | $\small {D} $-Lactic Acid Production by Sporolactobacillus inulinus Y2-8 Immobilized in Fibrous Bed Bioreactor Using Corn Flour Hydrolyzate | |
| Chu et al. | Dairy cow solid waste hydrolysis and hydrogen/methane productions by anaerobic digestion technology | |
| CN101402913B (en) | Method for producing propionic acid and co-producing vitamin B by batch fermentation method12Apparatus and process for | |
| EP4296366A1 (en) | Method for producing polyhydroxyalkanoate by fermentation of agricultural wastes | |
| CN107746862A (en) | Utilize the method for clostridium tyrobutyricum fermented maize skin hydrolyzate production butyric acid | |
| CN101544990A (en) | Method for producing gas fuel and byproduct cellulase by using biomass containing lignocellulose through fermentation | |
| CN102304480B (en) | Lactobacillus rhamnose strain for producing L-lactic acid efficiently and method for producing L-lactic acid by fermenting cassava and sugarcane molasses | |
| CN104357428A (en) | Liquid submerged fermentation method of xylanase | |
| CN101705261A (en) | Preparation method of Gamma-aminobutyric acid | |
| CN103952447B (en) | Method for producing succinic acid by fermentation under anaerobic condition | |
| CN109082370B (en) | Artificial rumen and use method thereof | |
| CN103243128A (en) | High-yield production method of GABA (gamma amino butyric acid) through mixed fermentation of brevibacterium tianjinese and lactobacillus plantarum | |
| CN103642680A (en) | Built-in disc fibrous bed reactor for fermenting production of butyric acid | |
| CN102041279B (en) | Biomembrane-electrodialysis coupling continuous production technology of L-lactic acid |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110831 Termination date: 20191120 |