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Applied and Environmental Microbiology, February 2003, p. 1263-1269, Vol. 69, No. 2
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.2.1263-1269.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Cloning and Analysis of a DNA Fragment Stimulating Avermectin Production in Various Streptomyces avermitilis Strains

Yong-Soon Hwang,1,{dagger} Eung-Soo Kim,2 Sándor Biró,3 and Cha-Yong Choi1*

School of Chemical Engineering, Seoul National University, Seoul,1 Division of Chemical Science and Biological Engineering, Inha University, Inchon, Korea,2 Department of Human Genetics, Medical and Health Science Center, University of Debrecen, H-4012 Debrecen, Hungary3

Received 24 January 2002/ Accepted 26 July 2002

To isolate a gene for stimulating avermectin production, a genomic library of Streptomyces avermitilis ATCC 31267 was constructed in Streptomyces lividans TK21 as the host strain. An 8.0-kb DNA fragment that significantly stimulated actinorhodin and undecylprodigiosin production was isolated. When wild-type S. avermitilis was transformed with the cloned fragment, avermectin production increased approximately 3.5-fold. The introduction of this fragment into high-producer (ATCC 31780) and semi-industrial (L-9) strains also resulted in an increase of avermectin production by more than 2.0- and 1.4-fold, respectively. Subclones were studied to locate the minimal region involved in stimulation of pigmented-antibiotic and avermectin production. An analysis of the nucleotide sequence of the entire DNA fragment identified eight complete and one incomplete open reading frame. All but one of the deduced proteins exhibited strong homology (68 to 84% identity) to the hypothetical proteins of Streptomyces coelicolor A3(2). The orfX gene product showed no significant similarity to any other protein in the databases, and an analysis of its sequence suggested that it was a putative membrane protein. Although the nature of the stimulatory effect is still unclear, the disruption of orfX revealed that this gene was intrinsically involved in the stimulation of avermectin production in S. avermitilis.

* Corresponding author. Mailing address: School of Chemical Engineering, College of Engineering, Seoul National University, Seoul 151-742, Korea. Phone: 82 2 880 7071. Fax: 82 2 874 1206. E-mail: choicy{at}snu.ac.kr.

{dagger} Present address: Samyang Genex Biotech Research Institute, Hwaam-dong, Yusung-gu, Taejeon, Korea.

Applied and Environmental Microbiology, February 2003, p. 1263-1269, Vol. 69, No. 2
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.2.1263-1269.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.





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