Microorganism resistant to threonine analogue and production of

Details Microorganism resistant to threonine analogue and production of Microorganism resistant to threonine analogue and production of

1997-02-19

2000-02-01

Achutamurthy, Ponnathapu

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing heterocyclic carbon compound having only o, n, s,...

435118, 43525233, 4352523, C12P 1718, C12P 1716, C12N 120

Patent

active

060201738

DESCRIPTION:

BRIEF SUMMARY
FIELD OF THE INVENTION

The present invention relates to a novel microorganism resistant to a threonine analogue and a process for producing biotin using the microorganism. The biotin obtainable by the invention can be used as a raw material for medicaments or cosmetics, feed additives, etc.


BACKGROUND OF THE INVENTION

Biotin (vitamin H) is a kind of vitamin B and is related to fatty acid synthesis or saccharide metabolism as a coenzyme of a carboxylase. About 10 tons of biotin has been produced by chemical synthesis processes every year for use as a raw material for medicaments or cosmetics, feed additives, etc. However, because these processes are complicated, biotin is very expensive. On the other hand, biotin production by fermentation processes has been studied for a long time. The fermentation processes have not become practical because their productivity is low.
Biotin production using gene manipulation techniques has been expected to provide inexpensive biotin. Some microorganisms modified by gene engineering techniques have been used for biotin production. For example, microorganisms belonging to the genus Escherichia such as a strain resistant to .alpha.-dehydrobiotin disclosed in e.g. JP-A 61-149091 are known as the modified microorganisms for the biotin production. Other known modified microorganisms for the biotin production include microorganisms belonging to the genus Bacillus modified by transforming Bacillus sphaericus and then providing resistance to thenoyltrifluoroacetone (JP-A 4-11894), microorganisms belonging to the genus Serratia modified by providing Serratia marcescens SB411 with ethionine-resistance followed by S-aminoethylcysteine-resistance and then transforming the resulting microorganism with a recombinant plasmid containing a biotin gene fragment (JP-A 5-199867), transformants of Serratia marcescens SB411 provided with resistance to actithiazic acid, a compound having biotin-like structure, or resistance to 5-(2-thienyl)-n-valeric acid (JP-A 2-27980), and transformants provided with resistance to a nicotinic acid analogue (Japanese Patent Application No. 6-311778).
However, the prior art processes for producing biotin are unsatisfactory for the industrial production of biotin. There is still a need for a process for producing biotin having increased biotin productivity.


OBJECTS OF THE INVENTION

The main object of the present invention is to provide a microorganism which can produce biotin in high yield.
Another object of the present invention is to provide a process for producing biotin using the above microorganism.
These objects as well as other objects and advantages of the present invention will become apparent to those skilled in the art from the following description with reference to the accompanying drawings.


SUMMARY OF THE INVENTION

S-Adenosylmethionine is essential for the main synthetic pathway from pimelyl CoA to desthiobiotin (a biotin precursor). The present inventors have expected that enhancing the biosynthetic pathway to methionine would enhance the supplying system of S-adenosylmethionine, thereby improving the accumulation of desthiobiotin and biotin.
In order to enhance the biosynthetic pathway to methionine, the present inventors have isolated a strain resistant to a threonine analogue from a biotin-producing microorganism, and obtained a strain with significantly increased desthiobiotin and biotin accumulation. After further studies based on this finding, the present invention has been accomplished.
The present invention provides a microorganism resistant to a threonine analogue, which has a plasmid containing part or whole of a biotin operon.
The present invention also provides a process for producing biotin, which comprises culturing a microorganism described above in a medium to produce and accumulate biotin in the medium, and collecting biotin.


BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a regulatory region of the biotin operon and a base sequence near the bio B initiation codon.
FIG. 2 is a restriction map of DNA of plasmid pXBA 312.

REFERENCES:
Max Eisenberg, "Regulation of the Biotin Operon in E. coli", Annals of the New York Academy of Sciences, vol. 447, pp. 335-349, 1985.

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