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Applied and Environmental Microbiology, December 2008, p. 7383-7390, Vol. 74, No. 23
0099-2240/08/$08.00+0     doi:10.1128/AEM.01759-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Knockout of the Erythromycin Biosynthetic Cluster Gene, eryBI, Blocks Isoflavone Glucoside Bioconversion during Erythromycin Fermentations in Aeromicrobium erythreum but Not in Saccharopolyspora erythraea{triangledown}

Andrew R. Reeves,{dagger} Ramya Seshadri,{ddagger} Igor A. Brikun,§ William H. Cernota, Melissa C. Gonzalez, and J. Mark Weber*

Fermalogic, Inc., 2201 West Campbell Park Drive, Chicago, Illinois 60612

Received 30 July 2008/ Accepted 25 September 2008

Isoflavone glucosides are valuable nutraceutical compounds and are present in commercial fermentations, such as the erythromycin fermentation, as constituents of the soy flour in the growth medium. The purpose of this study was to develop a method for recovery of the isoflavone glucosides as value-added coproducts at the end of either Saccharopolyspora erythraea or Aeromicrobium erythreum fermentation. Because the first step in isoflavone metabolism was known to be the conversion of isoflavone glucosides to aglycones by a β-glucosidase, we chose to knock out the only β-glucosidase gene known at the start of the study, eryBI, to see what effect this had on metabolism of isoflavone glucosides in each organism. In the unicellular erythromycin producer A. erythreum, knockout of eryBI was sufficient to block the conversion of isoflavone glucosides to aglycones. In S. erythraea, knockout of eryBI had no effect on this reaction, suggesting that other β-glucosidases are present. Erythromycin production was not significantly affected in either strain as a result of the eryBI knockout. This study showed that isoflavone metabolism could be blocked in A. erythreum by eryBI knockout but that eryBI knockout was not sufficient to block isoflavone metabolism in S. erythraea.

* Corresponding author. Mailing address: Fermalogic, Inc., 2201 West Campbell Park Drive, Chicago, IL 60612. Phone: (312) 738-0050. Fax: (312) 738-0963. E-mail: mark.weber{at}fermalogic.com

{triangledown} Published ahead of print on 3 October 2008.

{dagger} Present address: Coskata, Inc., 4575 Weaver Pkwy., Suite 100, Warrenville, IL 60555.

{ddagger} Present address: Abbott Molecular, 1300 East Touhy Avenue, Des Plaines, IL 60018.

§ Present address: Euclid Biosciences, 9800 Connecticut Drive, Crown Point, IN 46307.

Applied and Environmental Microbiology, December 2008, p. 7383-7390, Vol. 74, No. 23
0099-2240/08/$08.00+0     doi:10.1128/AEM.01759-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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