Transformant and its production process, and method for producing lactic acid
Provided is a transformant which can produce lactic acid with a high productivity without requiring neutralization with an alkali and is excellent in both of lactic acid production capability and growth ability and its production process, and a method for producing lactic acid by using the transformant. A transformant comprising 3 copies of a human lactate dehydrogenase gene that is introduced into a Schizosaccharomyces pombe host, wherein a gene encoding pyruvate decarboxylase 2 of the Schizosaccharomyces pombe host is deleted or inactivated.
1. A transformant comprising:
3 copies of a human lactate dehydrogenase gene that are introduced into a Schizosaccharomyces pombe host,
wherein a gene encoding pyruvate decarboxylase 2 of the Schizosaccharomyces pombe host is deleted or inactivated.
2. The transformant according to claim 1 ,
wherein the 3 copies of the human lactate dehydrogenase gene are integrated into Schizosaccharomyces pombe chromosome.
3. The transformant according to claim 2 ,
wherein each of the 3 copies of the human lactate dehydrogenase gene is introduced into a region that comprises from 10,000 bp upstream to 10,000 bp downstream of an eno101 gene locus, a region that comprises from 10,000 bp upstream to 10,000 bp downstream of a leu1 gene locus, and a region that comprises from 10,000 bp upstream to 10,000 bp downstream of a gpm1 gene locus.
4. A process for producing the transformant according to claim 1 , the process comprising:
introducing an expression cassette that comprises a promoter and a terminator capable of functioning in Schizosaccharomyces pombe and a human lactate dehydrogenase gene into 3 positions of chromosome of a Schizosaccharomyces pombe host to obtain the transformant; and
using a host in which a gene encoding pyruvate decarboxylase 2 is deleted or inactivated as the Schizosaccharomyces pombe host, or deleting or inactivating a gene encoding pyruvate decarboxylase 2 of the obtained transformant.
5. The process for producing a transformant according to claim 4 ,
wherein the expression cassette is introduced into a region selected from a region that comprises from 10,000 bp upstream to 10,000 bp downstream of an eno101 gene locus, a region that comprises from 10,000 bp upstream to 10,000 bp downstream of a leu1 gene locus, and a region that comprises from 10,000 bp upstream to 10,000 bp downstream of a gpm1 gene locus.
6. A method for producing lactic acid, comprising cultivating the transformant as defined in claim 1 , and recovering lactic acid from a culture broth.
7. The method for producing lactic acid according to claim 6 , wherein a culture broth having a glucose concentration of from 1 to 50 mass % is used for cultivating the transformant.
8. The method for producing lactic acid according to claim 6 , wherein the cultivating is further continued after pH of the culture broth becomes 3.5 or lower due to the lactic acid produced by the transformant.
9. The method for producing lactic acid according to claim 6 , wherein the cultivating is continued without neutralizing the lactic acid produced by the transformant in the culture broth.
10. The method for producing lactic acid according to claim 6 , wherein the lactic acid is separated from the culture broth without neutralizing the lactic acid produced by the transformant in the culture broth.