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Molecular genetics and enzymology of...

Grochowski, Laura L.

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Molecular genetics and enzymology of secondary metabolite biosynthesis: I. Isolation of natural product biosynthesis gene clusters from symbiotic marine organisms. II. Enzymology of blasticidin S biosynthesis.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Molecular genetics and enzymology of secondary metabolite biosynthesis: I. Isolation of natural product biosynthesis gene clusters from symbiotic marine organisms. II. Enzymology of blasticidin S biosynthesis./
Author:	Grochowski, Laura L.
Description:	229 p.
Notes:	Source: Dissertation Abstracts International, Volume: 65-07, Section: B, page: 3471.
Contained By:	Dissertation Abstracts International65-07B.
Subject:	Chemistry, Pharmaceutical. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3138458
ISBN:	0496856197

Molecular genetics and enzymology of secondary metabolite biosynthesis: I. Isolation of natural product biosynthesis gene clusters from symbiotic marine organisms. II. Enzymology of blasticidin S biosynthesis.
Grochowski, Laura L.

Molecular genetics and enzymology of secondary metabolite biosynthesis: I. Isolation of natural product biosynthesis gene clusters from symbiotic marine organisms. II. Enzymology of blasticidin S biosynthesis. - 229 p.

Source: Dissertation Abstracts International, Volume: 65-07, Section: B, page: 3471.

Thesis (Ph.D.)--Oregon State University, 2004.

Molecular genetic and enzymological techniques have been employed to study secondary metabolite biosynthesis. These investigations have focused on two projects: the cloning and heterologous expression of biosynthetic gene clusters from unculturable marine organisms and the characterization of individual enzymes involved in the biosynthesis of the antifungal agent blasticidin S.

ISBN: 0496856197Subjects--Topical Terms:

550957
Chemistry, Pharmaceutical.

Molecular genetics and enzymology of secondary metabolite biosynthesis: I. Isolation of natural product biosynthesis gene clusters from symbiotic marine organisms. II. Enzymology of blasticidin S biosynthesis.
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035 $a (UnM)AAI3138458
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100 1 $a Grochowski, Laura L. $3 1900854
245 1 0 $a Molecular genetics and enzymology of secondary metabolite biosynthesis: I. Isolation of natural product biosynthesis gene clusters from symbiotic marine organisms. II. Enzymology of blasticidin S biosynthesis.
300 $a 229 p.
500 $a Source: Dissertation Abstracts International, Volume: 65-07, Section: B, page: 3471.
500 $a Adviser: T. Mark Zabriskie.
502 $a Thesis (Ph.D.)--Oregon State University, 2004.
520 $a Molecular genetic and enzymological techniques have been employed to study secondary metabolite biosynthesis. These investigations have focused on two projects: the cloning and heterologous expression of biosynthetic gene clusters from unculturable marine organisms and the characterization of individual enzymes involved in the biosynthesis of the antifungal agent blasticidin S.
520 $a The marine environment is proving to be a valuable source of biologically active compounds, but problems associated with sustainable harvest, laboratory culture, and organic synthesis make obtaining sufficient quantities of compounds for drug development both difficult and expensive. A method has been developed for the isolation of biosynthetic gene clusters from complex marine microbe/invertebrate associations. Using this method a mixed polyketide synthase (PKS) and nonribosomal peptide synthetase (NRPS) gene cluster has been cloned from the marine sponge Jaspis splendens. The cloned gene cluster was found to code for a PKS with three extension modules and an NRPS with three extension modules. In addition, several open reading frames (ORFs) were identified that may be involved in the biosynthesis of the PKS starter molecule. Partial characterization of catalytic domains from the NRPS was also completed.
520 $a The second project centers on the characterization of enzymes involved in blasticidin S (BS) biosynthesis. Two ORFs were identified in the BS gene cluster encoding gene products predicted to be involved in the early steps of BS biosynthesis. The blsG gene product has sequence similarity to lysine 2,3-aminomutase and is believed to be involved in the formation of the beta-arginine moiety of BS. A series of heterologous expression studies were undertaken to determine the function of BlsG.
520 $a The product of blsM exhibits sequence homology with several nucleosidetransferases. blsM was cloned from the BS gene cluster, heterologously expressed in E. coli, and shown to catalyze the formation of cytosine using cytidine 5' -monophosphate as the preferred substrate. Point mutations were introduced in blsM to generate three BlsM mutant enzymes: S92D, E98A, and E98D. All three mutants lost cytidine 5'-monophosphate hydrolysis activity. Surprisingly, the BlsM S92D mutant exhibits cytidine deaminase activity when incubated with cytidine or deoxycytidine, resulting in the formation of uridine and deoxyuridine, respectively.
590 $a School code: 0172.
650 4 $a Chemistry, Pharmaceutical. $3 550957
650 4 $a Chemistry, Biochemistry. $3 1017722
650 4 $a Biology, Microbiology. $3 1017734
650 4 $a Biology, Molecular. $3 1017719
690 $a 0491
690 $a 0487
690 $a 0410
690 $a 0307
710 2 0 $a Oregon State University. $3 625720
773 0 $t Dissertation Abstracts International $g 65-07B.
790 1 0 $a Zabriskie, T. Mark, $e advisor
790 $a 0172
791 $a Ph.D.
792 $a 2004
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3138458