Cyclic lipopeptide acylase

Details Cyclic lipopeptide acylase Cyclic lipopeptide acylase

1998-12-31

2000-11-14

Lilling, Herbert J.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Hydrolase

435 681, 435212, 435228, 514 9, 514 11, 530317, 530318, C12N 950, C12N 986, C07K 512, C12P 2106

Patent

active

06146872&

DESCRIPTION:

BRIEF SUMMARY
TECHNICAL FIELD

This invention is concerned with an enzyme technology.
The present invention relates to a novel acylase which deacylates the acyl side chain of a cyclic lipopeptide compound and to a deacylation process comprising the use thereof.
More particularly, this invention relates to a novel acylase which deacylates the acyl side chain of FR901379 Substance, which is produced by Coleophoma sp. F-11899 (FERM BP-2635) (as described in Japanese Kokai Tokkyo Koho H3-184921), or any analog of FR901379 Substance and to a deacylation process using the same.


BACKGROUND ART

There has been a standing demand for an acylase capable of deacylating the acyl side chain of a cyclic lipopeptide compound, specifically said FR901379 Substance or an analog thereof, with good efficiency.


DISCLOSURE OF THE INVENTION

The inventors of this invention explored in earnest for a new acylase which might be able to deacylate the acyl side chain of a cyclic lipopeptide compound represented by FR901379 Substance, Echinocandin B and Aculeacin A, the latter two being analogs of FR901379 Substance. As a result, they discovered an acylase in the fermentation broth available upon culture of a certain filamentous fungus and succeeded in achieving the objective deacylation with effectiveness.
The characteristics of the above novel cyclic lipopeptide acylase and of the deacylation process using the enzyme are now described in detail.
The cyclic lipopeptide acylase-producing strain of the invention is first described. The filamentous fungus as a novel cyclic lipopeptide acylase producer specifically includes but is not limited to Oidiodendron sp. No. 30084, Oidiodendron echinulatum IFO 31963, Oidiodendron tenuissimum IFO 6798, Oidiodendron truncatum IFO 9951 and Oidiodendron truncatum IFO 31812, all of which belong to the genus Oidiodendron, and Verticillium sp. No. 30085 which belongs to the genus Verticillium.
The mycological characteristics of Oidiodendron sp. No. 30084 and Verticillium sp. No. 30085 are described below.
The fungus strain No. 30084 was isolated from a soil sample collected in Jouhoku-machi, Higashi Ibaraki-gun, Ibaraki Prefecture. This strain grew repressively on various media, forming colonies varying in color, e.g. greenish gray, brownish orange, yellowish white, etc., according to different kinds of culture media. On several media, strain No. 30084 formed anamorphs showing a conidial structure consisting of a dendritic conidiophore rising up from the surface of the medium and arthroconidia formed at its branches.
The detailed mycological characteristics of strain No. 30084 are as follows.
The cultural characteristics of this fungus on various agar media are summarized in Table 1. Colonies on malt extract agar grew repressively and spread to attain diameters from 1.5 to 2.0 cm after 2 weeks of incubation at 25.degree. C. The colony was circular and either raised as a whole or elevated peripherally and depressed in the center. The strain formed anamorphs in abundance, which presented with a powdery surface. The colony was greenish gray with a yellowish gray peripheral zone. The reverse side was light yellow with a yellowish white peripheral zone. On potato dextrose agar, too, the colony grew repressively and spread to attain diameters from 1.5 to 2.0 cm under the same cultural conditions as above. The surface of the colony was centrally elevated or convex, wrinkled, and somewhat felt-like, forming a small amount of anamorphs. The colony was brownish orange to light brown with an orange white peripheral zone. The reverse side was brown with an orange white peripheral zone.
The morphological characteristics of the strain was recorded by observing its growth on Miura's medium (Miura, K. and M. Kudo: Trans. Mycol. Soc. Japan, 11: 116-118, 1970). The conidiophore of strain No. 30084 stood erect from the surface of the medium and consisted of a tan-colored linear trunk and colorless intricate branches. This dendritic structure made the conidiophore clearly distinguishable from the vegetative hypha. The conidiophore wa

REFERENCES:
patent: H1638 (1997-03-01), Furuta et al.
patent: 5376634 (1994-12-01), Uwamoto et al.
patent: 5618787 (1997-04-01), Jamison et al.
patent: 5629289 (1994-12-01), Rodriguez
Computer Derwent Dgene Abstract 1997P-W45450 Peptide WO 9732975 New Cyclic Lipo:Peptide Acylase, Sep. 12, 1997.

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