WO2005071061A1 - A method for producing lactic acid with high concentration and high yield using lactic acid bacteria - Google Patents
A method for producing lactic acid with high concentration and high yield using lactic acid bacteria Download PDFInfo
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- WO2005071061A1 WO2005071061A1 PCT/KR2004/002991 KR2004002991W WO2005071061A1 WO 2005071061 A1 WO2005071061 A1 WO 2005071061A1 KR 2004002991 W KR2004002991 W KR 2004002991W WO 2005071061 A1 WO2005071061 A1 WO 2005071061A1
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- lactic acid
- producing
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- fermentation
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 title claims abstract description 119
- 235000014655 lactic acid Nutrition 0.000 title claims abstract description 59
- 239000004310 lactic acid Substances 0.000 title claims abstract description 59
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 24
- 241000894006 Bacteria Species 0.000 title description 8
- 241000186605 Lactobacillus paracasei Species 0.000 claims abstract description 9
- JVTAAEKCZFNVCJ-REOHCLBHSA-N L-lactic acid Chemical compound C[C@H](O)C(O)=O JVTAAEKCZFNVCJ-REOHCLBHSA-N 0.000 claims description 28
- 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 claims description 14
- 239000008103 glucose Substances 0.000 claims description 14
- 229930182843 D-Lactic acid Natural products 0.000 claims description 10
- JVTAAEKCZFNVCJ-UWTATZPHSA-N D-lactic acid Chemical compound C[C@@H](O)C(O)=O JVTAAEKCZFNVCJ-UWTATZPHSA-N 0.000 claims description 10
- 239000001888 Peptone Substances 0.000 claims description 10
- 108010080698 Peptones Proteins 0.000 claims description 10
- 229940022769 d- lactic acid Drugs 0.000 claims description 10
- 235000019319 peptone Nutrition 0.000 claims description 10
- 229940041514 candida albicans extract Drugs 0.000 claims description 9
- 239000012138 yeast extract Substances 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 6
- 240000008042 Zea mays Species 0.000 claims description 4
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 4
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 4
- 235000005822 corn Nutrition 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 230000012010 growth Effects 0.000 claims description 3
- 235000021109 kimchi Nutrition 0.000 abstract description 7
- 229920000747 poly(lactic acid) Polymers 0.000 abstract description 7
- 239000004626 polylactic acid Substances 0.000 abstract description 7
- 229920000704 biodegradable plastic Polymers 0.000 abstract description 5
- -1 chemistry Substances 0.000 abstract description 4
- 150000007524 organic acids Chemical class 0.000 abstract description 4
- 238000011109 contamination Methods 0.000 abstract description 3
- 239000002537 cosmetic Substances 0.000 abstract description 3
- 230000007613 environmental effect Effects 0.000 abstract description 3
- 239000004744 fabric Substances 0.000 abstract description 3
- 239000004615 ingredient Substances 0.000 abstract description 3
- 239000002535 acidifier Substances 0.000 abstract description 2
- 235000013409 condiments Nutrition 0.000 abstract description 2
- 238000004043 dyeing Methods 0.000 abstract description 2
- 235000013373 food additive Nutrition 0.000 abstract description 2
- 239000002778 food additive Substances 0.000 abstract description 2
- 235000019249 food preservative Nutrition 0.000 abstract description 2
- 239000005452 food preservative Substances 0.000 abstract description 2
- 229910052751 metal Inorganic materials 0.000 abstract description 2
- 239000002184 metal Substances 0.000 abstract description 2
- 150000002739 metals Chemical class 0.000 abstract description 2
- 239000004753 textile Substances 0.000 abstract description 2
- 238000000855 fermentation Methods 0.000 description 22
- 230000004151 fermentation Effects 0.000 description 22
- 239000002054 inoculum Substances 0.000 description 9
- 244000005700 microbiome Species 0.000 description 9
- 239000002609 medium Substances 0.000 description 7
- 241000186660 Lactobacillus Species 0.000 description 6
- 229940039696 lactobacillus Drugs 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 230000010261 cell growth Effects 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- LZCLXQDLBQLTDK-UHFFFAOYSA-N ethyl 2-hydroxypropanoate Chemical compound CCOC(=O)C(C)O LZCLXQDLBQLTDK-UHFFFAOYSA-N 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- SMZOUWXMTYCWNB-UHFFFAOYSA-N 2-(2-methoxy-5-methylphenyl)ethanamine Chemical compound COC1=CC=C(C)C=C1CCN SMZOUWXMTYCWNB-UHFFFAOYSA-N 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N 2-Propenoic acid Natural products OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000194033 Enterococcus Species 0.000 description 1
- 241000218587 Lactobacillus paracasei subsp. paracasei Species 0.000 description 1
- 241000183331 Lactobacillus paracasei subsp. tolerans Species 0.000 description 1
- 241000192132 Leuconostoc Species 0.000 description 1
- 241000235527 Rhizopus Species 0.000 description 1
- 241000194017 Streptococcus Species 0.000 description 1
- IKHGUXGNUITLKF-XPULMUKRSA-N acetaldehyde Chemical compound [14CH]([14CH3])=O IKHGUXGNUITLKF-XPULMUKRSA-N 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 229920006125 amorphous polymer Polymers 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229940116333 ethyl lactate Drugs 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 239000012770 industrial material Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000006872 mrs medium Substances 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 229920001451 polypropylene glycol Polymers 0.000 description 1
- 238000007639 printing Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/56—Lactic acid
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
Definitions
- the present invention relates to a method for producing lactic acid using novel lactic acid bacteria. More particularly, the present invention relates to a method for producing lactic acid with high concentration and high yield using
- Lactobacillus paracasei CJLA0310 KCCM-10542 that is separated and identified from Kimchi.
- Lactic acid is a very important organic acid and its applications are broad including food additive such as food preservative, condiment or acidifier, and industrial fields such as cosmetics, chemistry, metals, electronics, fabrics, dyeing textiles, and pharmaceutical industries.
- lactic acid is an essential ingredient of polylactic acid, one of biodegradable plastics.
- gas price increase oil depletion
- non- degradable petro-chemical based plastics These concerns turn people's interests to environmentally-friendly polymer materials against recalcitrant non-biodegradable plastics which are main causes of environmental contamination. Accordingly, demand for lactic acid has been increased markedly. Worldwide production of lactic acid is approximately 100,000 tons every year, posting about 5% of growth.
- lactic acid is a very reactive organic acid containing a hydroxyl group and a carboxylic group. It is also used as a key ingredient of many chemicals including polylactic acid, acetaldehyde, polypropylene glycol, acrylic acid, and 2,3-pentathione. Moreover, lactic acid is used for preparation of ethyl lactate that is a biodegradable and non-toxic solvent widely used in manufacture of electronic instruments, paints or fabrics, detergents, adhesives or printings. As described above, lactic acid is a very useful and valuable industrial material.
- lactic acid was produced through the traditional chemical synthesis or the biological fermentation process using carbohydrates as a substrate.
- the latter is preferred in many cases because the chemical synthesis creates other problems in addition to the material cost increase caused by the gas price increase or environmental contamination problems.
- the traditional chemical synthesis of lactic acid not only produces lactic acid but also an inactive D, L lactic acid in form of a racemic mixture that consists of equal amount of D- lactic acid and L-lactic acid.
- the composition ratio of the D-lactic acid and the L-lactic acid cannot be controlled. Therefore, when the racemic lactic acid is used for preparing polylactic acid an amorphous polymer with low fusing point is produced, which in turn becomes an obstacle to the broad range of applications.
- the biological fermentation process using microorganisms makes it possible to selectively produce D-lactic acid or L-lactic acid depending on the strain used.
- microorganisms such as Lactobacillus, Bacillus, Rhizopus, Streptococcus, and Enterococcus usually produce L-lactic acid.
- Microorganisms such as Leuconostoc and Lactobacillus vulgaricus usually produce D-lactic acid.
- the L-lactic acid is preferably used in cosmetic or food industries because the D-lactic acid is not metabolized in the body. Pure L-lactic acid can be used for the preparation of polylactic acid. In doing so, the polylactic acid is transformed into a crystal form having 180°C of fusion point.
- the L-lactic acid is applicable in replacement of various kinds of plastics.
- lactic acid has a great potential to be an environmentally-friendly replacement material in a wide range of industrial applications.
- it is very important to develop a method for producing lactic acid through the biological fermentation which is more economical than the traditional chemical synthesis. To this end, there is an urgent need to develop a lactic acid-producing strain and a cost-effective method for producing lactic acid.
- Fig. 1 is a graph showing the relation between fermentation time and cell concentration, glucose concentration and lactic acid concentration in lactic acid fermentation using Lactobacillus paracasei CJLA0310 in a culture medium consisting of 180g/L of glucose, 15gL of yeast extract, and lOg/L of peptone; and Fig.
- FIG. 2 is a graph showing the relation between fermentation time and cell concentration, glucose concentration and lactic acid concentration in lactic acid fermentation using Lactobacillus paracasei CJLA0310 in a culture medium consisting of 180g/L of glucose, 7g/L of peptone, and 15g/L of CSP(Corn Steep Powder).
- the present invention has been made in view of the above problems, and it is an object of the present invention to provide a lactic acid- producing microorganism to produce industrially useful lactic acid more effectively through biological fermentation. More specifically, the present invention is directed to develop a method for producing industrially useful lactic acid bacteria with high concentration and high yield, the method involving the separation of lactic acid bacteria characterized of excellent growth efficiency and lactic acid production capacity. It is another object of the present invention to provide a lactic acid producing strain that features high lactic acid yield during the biological lactic acid fermentation process, high lactic acid concentration in a fermentation broth at the end of the fermentation process, short fermentation time, high productivity and high economic efficiency.
- the above and other objects can be accomplished by the provision of a method for producing high- purity lactic acid using novel lactic acid bacteria separated and identified as Lactobacillus paracasei CJLA0310 KCCM-10542 featuring excellent lactic acid production capacity.
- a fermentation method for producing lactic acid with high concentration and high yield using a Lactobacillus paracasei CJLA0310 strain that is separated and identified from Kimchi and a method for adjusting a production ratio of L- lactic acid and D-lactic acid under proper culture conditions using the strain.
- Lactobacillus paracasei CJLA0310 is a lactic acid-producing strain separated from Kimchi, and under proper culture conditions it can produce an optically pure L-lactic acid with high concentration and high yield.
- the present invention provides biological production of the L-lactic acid which is one of industrially useful organic acids. Because the biological production makes it possible to mass-produce lactic acid with high yield and high productivity with respect to carbohydrates, the present invention can be very advantageously used for the food and environment industries.
- Example 1 Separation and identification of lactic acid producing bacteria The present inventors separated the fluid of Kimchi that was collected from a waste Kimchi disposal unit in Icheon 1 Kimchi factory of CJ CORP. located in
- the strain turned out to be a novel Lactobacillus microorganism exhibiting 100% of similarity to Lactobacillus paracasei subsp. paracasei and Lactobacillus paracasei subsp. tolerans. Therefore, the strain was named Lactobacillus paracasei CJLA0310.
- the strain was deposited with the KCCM (Korean Culture Center of Microorganisms) on December 4, 2003, and given the number KCCM-10542. [Table 1] 16s rDNA base sequence similarity between CJLA0310 and Lactobacillus standard strain
- Example 2 Lactic acid fermentation characteristics of novel microorganism (180g/L of glucose + 15g/L of yeast extract + lOg/L of peptone)
- 600ml of inoculum was first seeded for 18 hours in a medium consisting of 180g/L of glucose, 15g/L of yeast extract, and lOg/L of peptone.
- the inoculum was seeded in a fermenter (5L) containing 2.4L of the fermentation medium which consisted of 180g/L of glucose, 15g/L of yeast extract, and lOg/L of peptone.
- the inoculum was cultured for 50 hours at pH 6.0, 200rpm, and 37°C.
- Example 3 Lactic acid fermentation characteristics of novel microorganism (160g/L of glucose + 15g/L of yeast extract + 15g/L of peptone)
- 600ml of inoculum was first seeded for 18 hours in the medium of Example 2.
- the inoculum was seeded in a fermenter (5L) containing 2.4L of the fermentation medium which consisted of 160g/L of glucose, 15g/L of yeast extract, and 15g/L of peptone.
- the inoculum was cultured for 50 hours at pH 6.0, 200rpm, and 33, 35, 37, 39 and 41°C, respectively.
- the fermentation ratio of D-lactic acid and L- lactic acid could be adjusted according to the fermentation temperatures. [Table 2] Relation between production ratio of D-lactic acid and L-lactic acid and fermentation temperatures
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- Dairy Products (AREA)
Abstract
The present invention relates to a method for producing lactic acid with high concentration and high yield using Lactobacillus paracasei CJLA0310 KCCM-10542 that is separated and identified from Kimchi. Lactic acid is a very important organic acid with a wide range of applications including food additive such as food preservative, condiment or acidifier, and industrial fields such as cosmetics, chemistry, metals, electronics, fabrics, dyeing textiles, and pharmaceutical industries. Particularly, lactic acid is an essential ingredient of polylactic acid, one of biodegradable plastics to replace recalcitrant non-biodegradable plastics which are main causes of environmental contamination.
Description
A METHOD FOR PRODUCING LACTIC ACID WITH HIGH CONCENTRATION AND HIGH YIELD USING LACTIC ACID BACTERIA
[Technical Field] The present invention relates to a method for producing lactic acid using novel lactic acid bacteria. More particularly, the present invention relates to a method for producing lactic acid with high concentration and high yield using
Lactobacillus paracasei CJLA0310 KCCM-10542 that is separated and identified from Kimchi.
[Background Art] Lactic acid is a very important organic acid and its applications are broad including food additive such as food preservative, condiment or acidifier, and industrial fields such as cosmetics, chemistry, metals, electronics, fabrics, dyeing textiles, and pharmaceutical industries. In addition, lactic acid is an essential ingredient of polylactic acid, one of biodegradable plastics. In recent years, there have been several concerns such as gas price increase, oil depletion and non- degradable petro-chemical based plastics. These concerns turn people's interests to environmentally-friendly polymer materials against recalcitrant non-biodegradable plastics which are main causes of environmental contamination. Accordingly, demand for lactic acid has been increased markedly. Worldwide production of lactic acid is approximately 100,000 tons every year, posting about 5% of growth. Considering the gradual increase in the biodegradable plastics, the demand for lactic
acid will rise even higher within several years. Particularly, lactic acid is a very reactive organic acid containing a hydroxyl group and a carboxylic group. It is also used as a key ingredient of many chemicals including polylactic acid, acetaldehyde, polypropylene glycol, acrylic acid, and 2,3-pentathione. Moreover, lactic acid is used for preparation of ethyl lactate that is a biodegradable and non-toxic solvent widely used in manufacture of electronic instruments, paints or fabrics, detergents, adhesives or printings. As described above, lactic acid is a very useful and valuable industrial material. Typically, lactic acid was produced through the traditional chemical synthesis or the biological fermentation process using carbohydrates as a substrate. The latter is preferred in many cases because the chemical synthesis creates other problems in addition to the material cost increase caused by the gas price increase or environmental contamination problems. For example, the traditional chemical synthesis of lactic acid not only produces lactic acid but also an inactive D, L lactic acid in form of a racemic mixture that consists of equal amount of D- lactic acid and L-lactic acid. Unfortunately though, the composition ratio of the D-lactic acid and the L-lactic acid cannot be controlled. Therefore, when the racemic lactic acid is used for preparing polylactic acid an amorphous polymer with low fusing point is produced, which in turn becomes an obstacle to the broad range of applications. On the other hand, the biological fermentation process using microorganisms makes it possible to selectively produce D-lactic acid or L-lactic acid depending on the strain used. For example, microorganisms such as Lactobacillus, Bacillus, Rhizopus, Streptococcus, and Enterococcus usually produce L-lactic acid.
Microorganisms such as Leuconostoc and Lactobacillus vulgaricus usually produce D-lactic acid. The L-lactic acid is preferably used in cosmetic or food industries because the D-lactic acid is not metabolized in the body. Pure L-lactic acid can be used for the preparation of polylactic acid. In doing so, the polylactic acid is transformed into a crystal form having 180°C of fusion point. Also, physical properties of the polylactic acid can be controlled by adjusting the content of the D- lactic acid for the polymerization. Overall, the L-lactic acid is applicable in replacement of various kinds of plastics. As described above, lactic acid has a great potential to be an environmentally-friendly replacement material in a wide range of industrial applications. Thus, it is very important to develop a method for producing lactic acid through the biological fermentation which is more economical than the traditional chemical synthesis. To this end, there is an urgent need to develop a lactic acid-producing strain and a cost-effective method for producing lactic acid.
[Description of the Drawings] The above and other objects, features and other advantages of the present invention will be more clearly understood from the following detailed description taken in conjunction with the accompanying drawings, in which: Fig. 1 is a graph showing the relation between fermentation time and cell concentration, glucose concentration and lactic acid concentration in lactic acid fermentation using Lactobacillus paracasei CJLA0310 in a culture medium consisting of 180g/L of glucose, 15gL of yeast extract, and lOg/L of peptone; and
Fig. 2 is a graph showing the relation between fermentation time and cell concentration, glucose concentration and lactic acid concentration in lactic acid fermentation using Lactobacillus paracasei CJLA0310 in a culture medium consisting of 180g/L of glucose, 7g/L of peptone, and 15g/L of CSP(Corn Steep Powder).
[Disclosure] [Technical Problem] Therefore, the present invention has been made in view of the above problems, and it is an object of the present invention to provide a lactic acid- producing microorganism to produce industrially useful lactic acid more effectively through biological fermentation. More specifically, the present invention is directed to develop a method for producing industrially useful lactic acid bacteria with high concentration and high yield, the method involving the separation of lactic acid bacteria characterized of excellent growth efficiency and lactic acid production capacity. It is another object of the present invention to provide a lactic acid producing strain that features high lactic acid yield during the biological lactic acid fermentation process, high lactic acid concentration in a fermentation broth at the end of the fermentation process, short fermentation time, high productivity and high economic efficiency.
[Technical Solution]
In accordance with an aspect of the present invention, the above and other objects can be accomplished by the provision of a method for producing high- purity lactic acid using novel lactic acid bacteria separated and identified as Lactobacillus paracasei CJLA0310 KCCM-10542 featuring excellent lactic acid production capacity. In accordance with another aspect of the present invention, there is provided a fermentation method for producing lactic acid with high concentration and high yield using a Lactobacillus paracasei CJLA0310 strain that is separated and identified from Kimchi, and a method for adjusting a production ratio of L- lactic acid and D-lactic acid under proper culture conditions using the strain. In accordance with another aspect of the present invention, there is suggested the possibility of reducing production cost of lactic acid by utilizing low-price corn steep powder in replacement of yeast extract in a medium, which is known as a key factor for increasing the cost of manufacture. Although the preferred embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that various modifications, additions and substitutions are possible, without departing from the scope and spirit of the invention as disclosed in the accompanying claims.
[Advantageous Effects] Lactobacillus paracasei CJLA0310 is a lactic acid-producing strain separated from Kimchi, and under proper culture conditions it can produce an optically pure L-lactic acid with high concentration and high yield. The present
invention provides biological production of the L-lactic acid which is one of industrially useful organic acids. Because the biological production makes it possible to mass-produce lactic acid with high yield and high productivity with respect to carbohydrates, the present invention can be very advantageously used for the food and environment industries.
[Mode for carrying out the invention] Example 1: Separation and identification of lactic acid producing bacteria The present inventors separated the fluid of Kimchi that was collected from a waste Kimchi disposal unit in Icheon 1 Kimchi factory of CJ CORP. located in
Icheon-si, Gyeonggi-do, Korea. The fluid was then diluted with physiological saline solution (8.5g/L NaCl) and smeared uniformly over the MRS medium
(manufactured by Difco Co.). After two days of cultivation at 37°C, colonies were separated and particularly yellowish colonies among them were assumed to be lactic acid bacterium. 12 largest colonies out of the yellowish colonies were separated and cultivated, respectively, in the MRS liquid mediums at 37°C for two days, in order to obtain a strain of the highest cell growth rate and lactic acid production capacity. Part of base sequence (800bp) of 16S rDNA of the strain was compared with DNA base sequence database of 11 kinds of Lactobacillus standard strains. As shown in Table 1 below, it was observed that the DNA base sequence of the separated strain manifested 89% of similarity to that of the Lactobacillus standard strain. Especially, the strain turned out to be a novel Lactobacillus microorganism exhibiting 100% of similarity to Lactobacillus paracasei subsp. paracasei and
Lactobacillus paracasei subsp. tolerans. Therefore, the strain was named Lactobacillus paracasei CJLA0310. The strain was deposited with the KCCM (Korean Culture Center of Microorganisms) on December 4, 2003, and given the number KCCM-10542. [Table 1] 16s rDNA base sequence similarity between CJLA0310 and Lactobacillus standard strain
Example 2: Lactic acid fermentation characteristics of novel microorganism (180g/L of glucose + 15g/L of yeast extract + lOg/L of peptone) To cultivate the novel lactic acid CJLA0310, 600ml of inoculum was first seeded for 18 hours in a medium consisting of 180g/L of glucose, 15g/L of yeast extract, and lOg/L of peptone. Then, the inoculum was seeded in a fermenter (5L) containing 2.4L of the fermentation medium which consisted of 180g/L of glucose, 15g/L of yeast extract, and lOg/L of peptone. The inoculum was cultured for 50
hours at pH 6.0, 200rpm, and 37°C. As shown in Fig. 2, after 50 hours of the cultivation process 180g of glucose was consumed, the cell growth medium (O.D.600) = 24, and 181g/L of lactic acid was finally produced. Therefore, the lactic acid yield = 99.5%, and average volume productivity per hour = 3.85g/L/hr.
Example 3: Lactic acid fermentation characteristics of novel microorganism (160g/L of glucose + 15g/L of yeast extract + 15g/L of peptone) To cultivate the novel lactic acid CJLA0310, 600ml of inoculum was first seeded for 18 hours in the medium of Example 2. Then, the inoculum was seeded in a fermenter (5L) containing 2.4L of the fermentation medium which consisted of 160g/L of glucose, 15g/L of yeast extract, and 15g/L of peptone. The inoculum was cultured for 50 hours at pH 6.0, 200rpm, and 33, 35, 37, 39 and 41°C, respectively. As shown in Table. 2, the fermentation ratio of D-lactic acid and L- lactic acid could be adjusted according to the fermentation temperatures. [Table 2] Relation between production ratio of D-lactic acid and L-lactic acid and fermentation temperatures
Example 4: Lactic acid fermentation characteristics of novel microorganism (180g/L of glucose + 7g/L of peptone + 15g L of CSP) To cultivate the novel lactic acid CJLA0310, 600ml of inoculum was first
seeded for 18 hours in the medium of Example 2. Then, the inoculum was seeded in a fermenter (5L) containing 2.4L of the fermentation medium which consisted of 180g/L of glucose, 7g/L of peptone and 15g/L of CSP (corn steep powder). The inoculum was cultured for 70 hours at pH 6.0, 200rpm, and 37°C. As shown in Fig. 2, after 65 hours of the cultivation process 180g of glucose was consumed, the cell growth medium (O.D.600) = 24, and 179.1g/L of lactic acid was finally produced. Therefore, the lactic acid yield = 99.5%, and average volume productivity per hour = 2.76g/L/hr.
Claims
[1] A Lactobacillus paracasei CJLA0310 KCCM-10542 strain characterized of excellent lactic acid production capacity and high growth rate.
[2] A method for producing lactic acid with high concentration by cultivating the strain of claim 1 in a medium that comprises 160-180g/L of glucose, 15g/L of yeast extract, and 7-15g/L of peptone.
[3] The method according to claim 2, wherein the production ratio of D-lactic acid and L-lactic acid is adjusted by varying the cultivation temperature in a range of 33- 41°C.
[4] The method according to claim 2, wherein the medium comprises corn steep powder in replacement of yeast extract.
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| SI200431688T SI1702060T1 (en) | 2003-12-11 | 2004-11-18 | A method for producing lactic acid with high concentration and high yield using lactic acid bacteria |
| EP04821212A EP1702060B1 (en) | 2003-12-11 | 2004-11-18 | A method for producing lactic acid with high concentration and high yield using lactic acid bacteria |
| US10/582,389 US7682814B2 (en) | 2003-12-11 | 2004-11-18 | Method for producing lactic acid with high concentration and high yield using lactic acid bacteria |
| BRPI0417119-5A BRPI0417119A (en) | 2003-12-11 | 2004-11-18 | lactobacillus paracasei cjla0310 kccm10542 strain and method for producing high concentration lactic acid |
| PL04821212T PL1702060T3 (en) | 2003-12-11 | 2004-11-18 | A method for producing lactic acid with high concentration and high yield using lactic acid bacteria |
| DE602004031769T DE602004031769D1 (en) | 2003-12-11 | 2004-11-18 | PROCESS FOR THE PRODUCTION OF MILKYLIC ACID WITH HIGH CONCENTRATION AND HIGH EXPLOITATION USING MILK ACID BACTERIA |
| DK04821212.0T DK1702060T3 (en) | 2003-12-11 | 2004-11-18 | Process for the production of high concentration and high yield of lactic acid using lactic acid bacteria |
| AT04821212T ATE501245T1 (en) | 2003-12-11 | 2004-11-18 | METHOD FOR PRODUCING HIGH CONCENTRATION AND HIGH YIELD LACTIC ACID USING LACTIC ACID BACTERIA |
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| KR10-2003-0090204A KR100517065B1 (en) | 2003-12-11 | 2003-12-11 | Method for producing lactic acid with high concentration and high yield using lactic acid bacteria |
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Cited By (4)
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| US7326550B2 (en) | 1997-09-12 | 2008-02-05 | Tate & Lyle Ingredients Americas, Inc. | Yeast strains for the production of lactic acid |
| US7473540B2 (en) | 2005-09-22 | 2009-01-06 | Tate & Lyle Ingredients Americas, Inc. | Methods for selecting a yeast population for the production of an organic acid and producing an organic acid |
| WO2012036575A1 (en) * | 2010-09-16 | 2012-03-22 | Instytut Biotechnologii Przemysłu Rolno-Spożywczego | Method of production of fermented, pro-healthy fruit beverages |
| US8492127B2 (en) | 2010-05-20 | 2013-07-23 | Shanghai Jiao Tong University | Bacillus coagulans strains and their applications in L-lactic acid production |
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| CN100554405C (en) * | 2007-10-18 | 2009-10-28 | 中国科学院微生物研究所 | A kind of method and special-purpose lactobacillus rhamnosus thereof that produces L-lactic acid |
| KR101224854B1 (en) * | 2010-12-03 | 2013-01-23 | 대상 주식회사 | Mutants of Lactobacillus paracasei with Improved Glucose Tolerance and Acid Tolerance and their Uses |
| KR101106302B1 (en) * | 2011-04-19 | 2012-01-18 | 충청북도 청원군(청원군농업기술센터장) | Culturing method of lactobacillus plantarum and probiotic drinking water for live-stock by using it |
| US9795161B2 (en) | 2011-09-30 | 2017-10-24 | Riken Vitamin Co., Ltd. | Taste-improving agent |
| KR101355713B1 (en) * | 2011-11-17 | 2014-02-05 | 주식회사 아워홈 | Composition for cultural medium for delaying the over ripening of kimchi and preparation method for kimchi using the same |
| KR101347182B1 (en) * | 2011-12-16 | 2014-01-06 | 한국화학연구원 | Lactobacillus paracasei LA104 producing L-lactic aicd |
| KR101464656B1 (en) | 2012-06-15 | 2014-12-02 | 한국생명공학연구원 | Varient Microorganism Having Metabolites Producing Ability and Method for Preparing Metabolites Using the Same |
| US9222110B2 (en) | 2012-12-17 | 2015-12-29 | South Dakota Board Of Regents | Microorganism and method for lactic acid production |
| KR101584698B1 (en) * | 2015-04-29 | 2016-01-25 | 대상 주식회사 | Mutants of Lactobacillus paracasei Producing L-Lactic Acid with Improved Optical Purity and Their Uses |
| TWI577800B (en) * | 2015-10-29 | 2017-04-11 | 行政院原子能委員會核能研究所 | Enterococcus faecalis and uses of lactic acid production at high temperatures |
| CN105567787A (en) * | 2016-01-22 | 2016-05-11 | 光明乳业股份有限公司 | Method for quickly distinguishing lactic acid bacterium strains |
| DE102017101220B4 (en) | 2017-01-23 | 2019-03-21 | Thyssenkrupp Ag | Minimal medium for the fermentative conversion of mono- and / or disaccharides to lactic acid with Bacillus coagulans strains |
| CN109294940B (en) * | 2018-09-04 | 2021-06-29 | 湖南肯基因科技有限公司 | Lactobacillus zeae mutant strain and application thereof in high yield of lactic acid |
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| EP0861905A2 (en) * | 1997-02-27 | 1998-09-02 | Proge Farm S.r.l. | Novel lactobacilli strains useful in the treatment of disorders of the gastrointestinal system |
| WO1999029833A1 (en) * | 1997-12-08 | 1999-06-17 | Arlafoods Amba | Strain of bacteria of the species lactobacillus paracasei subsp. paracasei, composition thereof for use in food and product containing said strain |
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| Publication number | Publication date |
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| DK1702060T3 (en) | 2011-07-04 |
| CY1111557T1 (en) | 2015-10-07 |
| DE602004031769D1 (en) | 2011-04-21 |
| EP1702060B1 (en) | 2011-03-09 |
| CN1906290A (en) | 2007-01-31 |
| KR100517065B1 (en) | 2005-09-26 |
| EP1702060A1 (en) | 2006-09-20 |
| ES2363085T3 (en) | 2011-07-20 |
| SI1702060T1 (en) | 2011-09-30 |
| US20070117193A1 (en) | 2007-05-24 |
| KR20050057969A (en) | 2005-06-16 |
| US7682814B2 (en) | 2010-03-23 |
| PL1702060T3 (en) | 2011-10-31 |
| ATE501245T1 (en) | 2011-03-15 |
| EP1702060A4 (en) | 2007-03-21 |
| BRPI0417119A (en) | 2007-03-06 |
| CN100417718C (en) | 2008-09-10 |
| PT1702060E (en) | 2011-06-28 |
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