Bacillus strains which produce HMG-CoA reductase inhibitors

Details Bacillus strains which produce HMG-CoA reductase inhibitors Bacillus strains which produce HMG-CoA reductase inhibitors

2001-04-10

2002-10-22

Lilling, Herbert J. (Department: 1651)

Chemistry: molecular biology and microbiology

Micro-organism, per se ; compositions thereof; proces of...

Bacteria or actinomycetales; media therefor

C435S125000, C435S135000, C435S136000

Reexamination Certificate

active

06468780

ABSTRACT:

TECHNICAL FIELD
The present invention relates to a process for producing compounds which inhibit hydroxymethylglutaryl-coenzyme A reductase (hereinafter abbreviated as HMG-COA reductase) and have the activity to lower the serum cholesterol level, etc.
BACKGROUND ART
It is known that a compound represented by general formula (VI-a):
(wherein R
1
represents a hydrogen atom or an alkali metal) [hereinafter referred to as Compound (VI-a)] or the lactone form of Compound (VI-a) represented by general formula (VI-b):
[hereinafter referred to as Compound (VI-b)] inhibits HMG-COA reductase and exhibits the activity to lower the serum cholesterol level, etc. [The Journal of Antibiotics, 29, 1346 (1976)].
Some microorganisms are known to have the ability to convert a compound represented by general formula (V-a):
(wherein R
1
represents a hydrogen atom or an alkali metal) [hereinafter referred to as Compound (V-a)] or the lactone form of Compound (V-a) represented by general formula (V-b):
[hereinafter referred to as Compound (V-b)] into Compound (VI-a) or Compound (VI-b). Such microorganisms include those belonging to the genus Absidia, Cunninghamella, Syncephalasporum or Streptomyces (Japanese Published Unexamined Patent Application No. 50894/82), those belonging to the genus Actinomucor, Circinella, Gongronella, Mortierella, Mucor, Phycomyces, Rhyzopus, Syncephalastrum, Zygorhynchus, Pycnoporus, Rhizoctonia or Nocardia [The Journal of Antibiotics, 36, 887 (1983)], those belonging to the genus Amycolata, Saccharopolyspora, Amycolatopsis or Saccharothrix (Japanese Published Unexamined Patent Application No. 184670/95) and those belonging to the genus Actinomadura (WO96/40863).
The above microorganisms belong to actinomycetes or filamentous fungi. So far, there has not been known a microorganism which belongs to bacteria and has the ability to convert Compound (V-a) or Compound (V-b) into Compound (VI-a) or Compound (VI-b), respectively, like those of the present invention. Actinomycetes and filamentous fungi have the drawback that their growth rate is lower than that of bacteria and thus more time is required for obtaining enough cells for the reaction. Further, there is the problem of controlling the culturing of actinomycetes and filamentous fungi in a fermenter. As actinomycetes and filamentous fungi grow by elongating hyphae, the viscosity of the culture rises as they grow in a fermenter. This often causes shortage of oxygen and makes the culture unhomogenous, which will lower the efficiency of reaction. To solve this problem of oxygen shortage and keep the culture homogenous, the stirring rate of the fermenter must be raised; but hyphae are liable to be cut by stirring at a higher rate, which will lower the activity of microorganisms [Fundamentals of Fermentation Technology, p. 169-190,P.F. Stansbury, A. Whitakaer, Gakkai Shuppan Center (1988)]. Culturing of actinomycetes and filamentous fungi involves such problems. On the other hand, culturing of bacteria, which do not form hyphae, can be readily carried out because the viscosity of the culture hardly rises, and insufficiency of aeration and lack of homogeneity of the culture are seldom observed.
In the DNA recombination technology, expression of genes in bacteria such as
Escherichia coli
is commonly carried out. However, it is generally difficult to efficiently express genes of actinomycetes and filamentous fungi because their codon usage are widely different from those of bacteria such as
Escherichia coli.
The available tools for efficient expression of genes in actinomycetes, such as vectors and promoters are limited. Therefore, it is desirable to employ bacteria, in which various vectors, promoters, etc. can be used, in order to express genes at a high level and to carry out reactions more efficiently. Any genes from bacteriacan bereadily expressed in bacteria at a high level.
DISCLOSURE OF THE INVENTION
An object of the present invention is to provide a process for producing a compound which inhibits HMG-COA reductase and has the activity to lower the serum cholesterol level, etc.
The present invention relates to a process for producing a compound represented by general formula (II-a):
(wherein R
1
represents a hydrogen atom, a substituted or unsubstituted alkyl group, or an alkali metal; and R
2
represents a substituted or unsubstituted alkyl or aryl group) [hereinafter referred to as Compound (II-a)] or the lactone form of Compound (II-a) represented by general formula (II-b):
(wherein R
2
represents a substituted or unsubstituted alkyl or aryl group) [hereinafter referred to as Compound (II-b)] which comprises subjecting a compound represented by general formula (I-a):
(wherein R
1
represents a hydrogen atom, a substituted or unsubstituted alkyl group, or an alkali metal; and R
2
represents a substituted or unsubstituted alkyl or aryl group) [hereinafter referred to as Compound (I-a)] or the lactone form of Compound (I-a) represented by general formula (I-b):
(wherein R
2
represents a substituted or unsubstituted alkyl or aryl group) [hereinafter referred to as Compound (I-b)] to the action of an enzyme source derived from a microorganism belonging to the genus Bacillus and capable of converting Compound (I-a) or Compound (I-b) into Compound (II-a) or Compound (II-b) in a reaction mixture to form Compound (II-a) or Compound (II-b) in the reaction mixture, and recovering Compound (II-a) or Compound (II-b) from the reaction mixture.
Examples of the alkyl groups include straight-chain or branched alkyl groups having 1-10 carbon atoms, preferably 1-6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, neopentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl and decyl, and branched-chain isomers thereof.
Examples of the aryl group include phenyl and naphthyl.
Examples of the substituent of the substituted alkyl group include halogen, hydroxy, amino, alkoxy, and aryl
Examples of the substituent of the substituted aryl group include halogen, hydroxy, amino, alkyl, and alkoxy.
The alkyl moiety of the alkoxy has the same significance as the alkyl group defined above.
The alkali metal means the elements of lithium, sodium, potassium, rubidium, cesium and francium.
Any enzyme source may be used in the present invention as long as it is derived from a microorganism belonging to the genus Bacillus and it has the activity to convert Compound (I-a) or Compound (I-b) into Compound (II-a) orCompound (II-b), respectively. Enzyme sources useful in the invention include microorganisms belonging to the genes Bacillus and having the activity to convert Compound (I-a) or Compound (I-b) into Compound (II-a) or Compound (II-b), cultures or cells of said microorganisms, treated matters thereof, and enzymes extracted from said microorganisms.
Examples of the microorganisms belonging to the genus Bacillus and having the activity to convert Compound (I-a) or Compound (I-b) into Compound (II-a) or Compound (II-b) are those belonging to
Bacillus laterosporus, Bacillus badius, Bacillus brevis, Bacillus alvei, Bacillus circulans, Bacillus macerans, Bacillus megaterium, Bacillus pumilus
and
Bacillus subtilis.
More specific examples thereof are
Bacillus laterosporus
ATCC 4517
, Bacillus badius
ATCC 14574
, Bacillus brevis
NRRL B-8029, Bacillus sp. PV-6,Bacillus sp. PV-7
, Bacillus alvei
ATCC 6344
, Bacillus circulans
NTCT-2610
, Bacillus macerans NCIB-
9368
,Bacillus megaterium
ATCC 10778
, Bacillus megateriumATCC
11562
,Bacillus megaterium
ATCC 13402
, Bacillus megateriumATCC
15177
,Bacillus mecaterium
ATCC 15450
, Bacillus megateriumATCC
19213
,Bacillus megaterium
IAM 1032
, Bacillus pumilus FERM BP-
2064
and Bacillus subtilis
ATCC 6051.
Also useful are subcultures, mutants or derivatives of the above microorganisms and recombinants prepared by recombinant DNA techniques. Bacillus sp. PV-6 and Bacillus sp. PV-7 were newl

Affiliated with

Also associated with

Rating

Bacillus strains which produce HMG-CoA reductase inhibitors does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Bacillus strains which produce HMG-CoA reductase inhibitors, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Bacillus strains which produce HMG-CoA reductase inhibitors will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2926780