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Production of bioarailable folic acid

A technology of microorganisms and monoglutamyl folate, which is applied in the field of various raw and microbial foods, can solve problems such as which specific gene is not used or which result is expected.

Inactive Publication Date: 2004-07-14
CAMPINA BV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It also suggests transforming these coryneform bacteria genes into coryneform bacteria or other microorganisms, but it doesn't have any protocol as to which specific gene to use or what outcome to expect

Method used

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  • Production of bioarailable folic acid
  • Production of bioarailable folic acid
  • Production of bioarailable folic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0024] Example 2 illustrates the overexpression of GTP-cyclohydrolase (EC 3.5.4.16) and 2-amino-4-hydroxy-6-hydroxymethyldihydropterin pyrophosphatase (EC 2.7.6) in Lactococcus lactis .3) the role of. In Lactococcus lactis, the gene gch encodes a bifunctional protein with these two enzymes. This effect not only increased total folic acid production, but also specifically increased monoglutamyl folate production and decreased polyglutamyl folate production in the medium using recombinant Lactococcus lactis. This can be explained by the relatively low activity of folyl polyglutamate synthase, encoded by the gene folC, which is not sufficiently active to accommodate the higher levels of folate produced by overexpression of GTP-cyclohydrolase. Biosynthesis. Monoglutamyl folic acid is more easily secreted by Lactococcus lactis, so that the fermentation medium not only has an increased level of folic acid, but also has a more bioavailable form that is secreted into the medium in t...

Embodiment 1

[0028] Materials and methods

[0029] The mature endopeptidase human γ-glutamyl hydrolase was obtained from the full-length cDNA cloned in the vector pCR2 (Yao et al., 1996a) by polymerase chain reaction. Vectors were provided by the Molecular Diagnostics Laboratory, Wadsworth Center, Albany, New York. The following primers were used to obtain an 885 bp PCR product encoding mature human γ-glutamyl hydrolase:

[0030] HGH-f(CATGCCATGGGACCCCACGGCGACACCGCCAAG) and

[0031] HGH-r (GCTCTAGATCAATCAAATATGTAACATTGCTG).

[0032] The forward primer was extended at the 5' end to create an NcoI restriction site enabling a transcriptional fusion to the nisin-inducible promoter of vector pNZ8048 (Kuipers et al.). Use of the forward primer as described results in slight modification of the mature gene (nucleotides in italics). The reverse primer was extended at the 3' end to create an XbaI restriction site that allowed cohesive end ligation to vector pNZ8048. A newly synthesized vector ...

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Abstract

The invention provides a process of producing bio-available folate, i.e. folic acid having an increased proportion of monoglutamyl folate and a decreased proportion of polyglutamyl folate, by culturing micro-organisms containing an active heterologous or homologous polyglutamyl hydrolase activity or containing increased activities of folate biosynthesis enzymes. The genes encoding the polyglutamyl hydrolase and the folate biosynthesis enzymes may be of various origin, e.g. from rodents or other mammals including man. Also provided is a foodstuff, especially a dairy product containing such monoglutamyl folate-producing micro-organisms.

Description

[0001] The present invention relates to the use of genetically modified microorganisms to increase the overall production of folic acid, and in particular the production of bioavailable folic acid, ie, folic acid that is readily absorbed by the gastrointestinal tract of mammals. Background technique [0002] Folic acid (N-[4-{[(2-amino-1,4-dihydro-4-oxo-pterinyl)methyl]amino}benzoyl]-L-glutamic acid; Vitamin B 11 ; Vitamin M) is a vitamin that promotes hematopoiesis, and it is gradually being recognized that it plays an important role in the health and wound healing of humans and animals. Mammals cannot produce folic acid. The main sources of folic acid are plants, especially spinach and other green leafy vegetables, grasses, yeast and other microorganisms, and indirectly, animal organs (kidneys, livers). At present, folic acid is supplemented by injection etc. or vitamin preparations. However, injecting these vitamins outside the regular diet is expensive and not always acc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23C9/123A23C19/032A23C21/02A23L1/30A23L1/305A23L29/00A23L33/00A23L33/15A61K31/525A61K35/66A61K35/74A61P3/02A61P7/06C12N1/00C12N1/20C12N1/21C12N9/48C12N15/09C12N15/52C12P7/44C12P13/02C12P17/12C12P17/18C12R1/01
CPCA23L1/0345A23C2220/202C12P17/182A23L1/302A23V2002/00A61K38/50C12N9/1235C12N9/48C12R1/00C12N9/78C12N5/10A61K38/45A61K36/06C12N15/52A23L29/065A23L33/15A61P3/02A61P7/06A23V2300/21
Inventor 威尔伯特·费克·亨里克斯·西比斯马杰罗恩·休根霍尔兹伊格尔·米劳玛丽亚·约翰娜·凯瑟琳娜·斯塔瑞恩布尔格米希尔·克勒瑞比赞姆威廉.迈因德特.德-福斯
Owner CAMPINA BV
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