A study of iterative type II polyketide synthases, using bacterial genes cloned from soil DNA: a means to access and use genes from uncultured microorganisms

被引:99
作者
Seow, KT
Meurer, G
Gerlitz, M
WendtPienkowski, E
Hutchinson, CR
Davies, J
机构
[1] UNIV WISCONSIN,SCH PHARM,MADISON,WI 53706
[2] TERRAGEN DIVERS INC,VANCOUVER,BC V6T 1Z3,CANADA
[3] UNIV WISCONSIN,DEPT BACTERIOL,MADISON,WI 53706
[4] UNIV BRITISH COLUMBIA,WE CTR MICROBIAL DIVERS,VANCOUVER,BC V5Z 1M9,CANADA
关键词
D O I
10.1128/jb.179.23.7360-7368.1997
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
To examine as randomly as possible the role of the beta-ketoacyl and acyl carrier protein (ACP) components of bacterial type II polyketide synthases (PKSs), homologs of the chain-length-factor (CLF) genes were cloned from the environmental community of microorganisms. With PCR primers derived from conserved regions of known ketosynthase (KSalpha) and ACP genes specifying the formation of 16- to 24-carbon polyketides, two CLF (KSbeta) genes were cloned from unclassified streptomycetes isolated from the soil, and two were cloned from soil DNA without the prior isolation of the parent microorganism. The sequence and deduced product of each gene were distinct from those of known KSbeta genes and, by phylogenetic analysis, belonged to antibiotic-producing PKS gene clusters. Hybrid PKS gene cassettes were constructed with each novel KSbeta gene substituted for the actI-ORF2 or tcmL KSbeta subunit genes, along with the respective actI-ORF1 or tcmK KSalpha, tcmM ACP, and tcmN cyclase genes, and were found to produce an octaketide or decaketide product characteristic of the ones known to be made by the heterologous KSalpha gene partner. Since substantially less than 1% of the microorganisms present in soil are thought to be cultivatable by standard methods, this work demonstrates a potential way to gain access to a more extensive range of microbial molecular diversity and to biosynthetic pathways whose products can be tested for biological applications.
引用
收藏
页码:7360 / 7368
页数:9
相关论文
共 74 条
[1]   CHARACTERIZATION OF ACTI-HOMOLOGOUS DNA ENCODING POLYKETIDE SYNTHASE GENES FROM THE MONENSIN PRODUCER STREPTOMYCES-CINNAMONENSIS [J].
ARROWSMITH, TJ ;
MALPARTIDA, F ;
SHERMAN, DH ;
BIRCH, A ;
HOPWOOD, DA ;
ROBINSON, JA .
MOLECULAR & GENERAL GENETICS, 1992, 234 (02) :254-264
[2]  
ATLAS RM, 1982, EXPT MICROBIAL ECOLO, P84
[3]   THE MULTIFUNCTIONAL 6-METHYLSALICYLIC ACID SYNTHASE GENE OF PENICILLIUM-PATULUM - ITS GENE STRUCTURE RELATIVE TO THAT OF OTHER POLYKETIDE SYNTHASES [J].
BECK, J ;
RIPKA, S ;
SIEGNER, A ;
SCHILTZ, E ;
SCHWEIZER, E .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1990, 192 (02) :487-498
[4]   ANALYSIS OF A POLYKETIDE SYNTHESIS-ENCODING GENE-CLUSTER OF STREPTOMYCES-CURACOI [J].
BERGH, S ;
UHLEN, M .
GENE, 1992, 117 (01) :131-136
[5]   ANALYSIS OF THE NUCLEOTIDE-SEQUENCE OF THE STREPTOMYCES-GLAUCESCENS TCML GENES PROVIDES KEY INFORMATION ABOUT THE ENZYMOLOGY OF POLYKETIDE ANTIBIOTIC BIOSYNTHESIS [J].
BIBB, MJ ;
BIRO, S ;
MOTAMEDI, H ;
COLLINS, JF ;
HUTCHINSON, CR .
EMBO JOURNAL, 1989, 8 (09) :2727-2736
[6]   CLONING, SEQUENCING AND DEDUCED FUNCTIONS OF A CLUSTER OF STREPTOMYCES GENES PROBABLY ENCODING BIOSYNTHESIS OF THE POLYKETIDE ANTIBIOTIC FRENOLICIN [J].
BIBB, MJ ;
SHERMAN, DH ;
OMURA, S ;
HOPWOOD, DA .
GENE, 1994, 142 (01) :31-39
[7]   A HYDROXYLASE-LIKE GENE-PRODUCT CONTRIBUTES TO SYNTHESIS OF A POLYKETIDE SPORE PIGMENT IN STREPTOMYCES-HALSTEDII [J].
BLANCO, G ;
PEREDA, A ;
BRIAN, P ;
MENDEZ, C ;
CHATER, KF ;
SALAS, JA .
JOURNAL OF BACTERIOLOGY, 1993, 175 (24) :8043-8048
[8]   GENETIC COMPLEMENTATION OF STREPTOMYCES-TENDAE DEFICIENT IN NIKKOMYCIN PRODUCTION [J].
BORMANN, C ;
ABERLE, K ;
FIEDLER, HP ;
SCHREMPF, H .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 1990, 32 (04) :424-430
[9]   Twenty-five coregulated transcripts define a sterigmatocystin gene cluster in Aspergillus nidulans [J].
Brown, DW ;
Yu, JH ;
Kelkar, HS ;
Fernandes, M ;
Nesbitt, TC ;
Keller, NP ;
Adams, TH ;
Leonard, TJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (04) :1418-1422
[10]   BIOSYNTHESIS OF FREDERICAMYCIN-A, A NEW ANTITUMOR ANTIBIOTIC [J].
BYRNE, KM ;
HILTON, BD ;
WHITE, RJ ;
MISRA, R ;
PANDEY, RC .
BIOCHEMISTRY, 1985, 24 (02) :478-486