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The standalone aminopeptidase PepN catalyzes the maturation of blasticidin S from leucylblasticidin S.

Yu G, Li L, Liu X, Liu G, Deng Z, Zabriskie MT, Jiang M, He X - Sci Rep (2015)

Bottom Line: PepN1 and PepN2, two neighboring PepN homologs from Streptomyces lividans were purified in E. coli but displayed ca.100-fold difference in LBS hydrolytic activity.Overexpression of pepN1 in WJ2 enhanced BS yield by 100% and lowered the ratio of LBS to BS from 2:1 to 2:3.This work presents the expansion of the biological role for PepN in antibiotic maturation and the first report of hydrolysis of beta amide linkage by this conserved enzyme.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Microbial Metabolism and School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai 200030 (China).

ABSTRACT
The peptidyl nucleoside blasticidin S (BS) isolated from Streptomyces griseochromogenes was the first non-mercurial fungicide used on a large scale to prevent rice blast. In the biosynthesis of BS, leucylblasticidin S (LBS) was suggested as the penultimate metabolite with 20-fold less inhibitory activity than the final product BS. Incomplete conversion of LBS to BS at a variable efficiency ranging from 10% to 90% was observed either in the native strain S. griseochromogenes or a heterologous producer Streptomyces lividans WJ2. In this study, we determined that maturation of BS from LBS is not a spontaneous process but is governed by a standalone peptidase PepN, which hydrolyzes LBS in a pH-sensitive way with most appropriate of pH 7~8 but is inactive when the pH is below 5 or above 10. PepN1 and PepN2, two neighboring PepN homologs from Streptomyces lividans were purified in E. coli but displayed ca.100-fold difference in LBS hydrolytic activity. Overexpression of pepN1 in WJ2 enhanced BS yield by 100% and lowered the ratio of LBS to BS from 2:1 to 2:3. This work presents the expansion of the biological role for PepN in antibiotic maturation and the first report of hydrolysis of beta amide linkage by this conserved enzyme.

No MeSH data available.


Related in: MedlinePlus

The yield of BS and LBS in S. lividans YGY7 (WJ2::pIB139), S. lividans YGY6 (WJ2::pIB139 + pepN1).The ratio of BS to LBS yield in the YGY6 is 3:2 as compared to that of 1:2 in YGY7, the yield of BS has nearly doubled.
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f6: The yield of BS and LBS in S. lividans YGY7 (WJ2::pIB139), S. lividans YGY6 (WJ2::pIB139 + pepN1).The ratio of BS to LBS yield in the YGY6 is 3:2 as compared to that of 1:2 in YGY7, the yield of BS has nearly doubled.

Mentions: As LBS is 20-fold less active than the final product BS23, in vivo conversion of LBS to BS by overexpression of pepN1 in S. lividans WJ2 is desired. pepN1 under the control of constitutive expression promoter PermE* were constructed and introduced into WJ2 to generate mutant strains S. lividans YGY6. The ratio of BS to LBS yield in the YGY6 is 3:2 as compared to that of 1:2 in WJ2::pIB139, the absolute yield of BS increased 100% upon the overexpression of PepN1(Fig. 6).These results demonstrated that overexpression of pepN1 is an efficient way to enhance BS production.


The standalone aminopeptidase PepN catalyzes the maturation of blasticidin S from leucylblasticidin S.

Yu G, Li L, Liu X, Liu G, Deng Z, Zabriskie MT, Jiang M, He X - Sci Rep (2015)

The yield of BS and LBS in S. lividans YGY7 (WJ2::pIB139), S. lividans YGY6 (WJ2::pIB139 + pepN1).The ratio of BS to LBS yield in the YGY6 is 3:2 as compared to that of 1:2 in YGY7, the yield of BS has nearly doubled.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664946&req=5

f6: The yield of BS and LBS in S. lividans YGY7 (WJ2::pIB139), S. lividans YGY6 (WJ2::pIB139 + pepN1).The ratio of BS to LBS yield in the YGY6 is 3:2 as compared to that of 1:2 in YGY7, the yield of BS has nearly doubled.
Mentions: As LBS is 20-fold less active than the final product BS23, in vivo conversion of LBS to BS by overexpression of pepN1 in S. lividans WJ2 is desired. pepN1 under the control of constitutive expression promoter PermE* were constructed and introduced into WJ2 to generate mutant strains S. lividans YGY6. The ratio of BS to LBS yield in the YGY6 is 3:2 as compared to that of 1:2 in WJ2::pIB139, the absolute yield of BS increased 100% upon the overexpression of PepN1(Fig. 6).These results demonstrated that overexpression of pepN1 is an efficient way to enhance BS production.

Bottom Line: PepN1 and PepN2, two neighboring PepN homologs from Streptomyces lividans were purified in E. coli but displayed ca.100-fold difference in LBS hydrolytic activity.Overexpression of pepN1 in WJ2 enhanced BS yield by 100% and lowered the ratio of LBS to BS from 2:1 to 2:3.This work presents the expansion of the biological role for PepN in antibiotic maturation and the first report of hydrolysis of beta amide linkage by this conserved enzyme.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Microbial Metabolism and School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai 200030 (China).

ABSTRACT
The peptidyl nucleoside blasticidin S (BS) isolated from Streptomyces griseochromogenes was the first non-mercurial fungicide used on a large scale to prevent rice blast. In the biosynthesis of BS, leucylblasticidin S (LBS) was suggested as the penultimate metabolite with 20-fold less inhibitory activity than the final product BS. Incomplete conversion of LBS to BS at a variable efficiency ranging from 10% to 90% was observed either in the native strain S. griseochromogenes or a heterologous producer Streptomyces lividans WJ2. In this study, we determined that maturation of BS from LBS is not a spontaneous process but is governed by a standalone peptidase PepN, which hydrolyzes LBS in a pH-sensitive way with most appropriate of pH 7~8 but is inactive when the pH is below 5 or above 10. PepN1 and PepN2, two neighboring PepN homologs from Streptomyces lividans were purified in E. coli but displayed ca.100-fold difference in LBS hydrolytic activity. Overexpression of pepN1 in WJ2 enhanced BS yield by 100% and lowered the ratio of LBS to BS from 2:1 to 2:3. This work presents the expansion of the biological role for PepN in antibiotic maturation and the first report of hydrolysis of beta amide linkage by this conserved enzyme.

No MeSH data available.


Related in: MedlinePlus