TAILIEUCHUNG - Kato and Iefuji AMB Express 2011, 1:7 http://www.amb-express.com/content/1/1/7 ORIGINAL Open

Kato and Iefuji AMB Express 2011, 1:7 ORIGINAL Open Access Breeding of a new wastewater treatment yeast by genetic engineering Miyoshi Kato1 and Haruyuki Iefuji1,2* Abstract We previously developed a host vector system for the wastewater treatment yeast Hansenula fabianii J640. The promoter and terminator regions of the gene encoding glucoamylase from H. fabianii J640 were used for a new expression vector, pHFGE-1. The performance of pHFGE-1 was compared with that of the widely used pG-1 transformant vector. H. fabianii J640 (HF-TAMY) cells were transformed with pHFGE-1, and Saccharomyces cerevisiae YPH-499 (SC-TAMY) cells were transformed with pG-1, both of which carried the Taka-amylase. Expression. | o AMB Express a SpringerOpen Journal Kato and lefuji AMB Express 2011 1 7 http content 1 1 7 ORIGINAL Open Access Breeding of a new wastewater treatment yeast by genetic engineering Miyoshi Kato1 and Haruyuki lefuji1 2 Abstract We previously developed a host vector system for the wastewater treatment yeast Hansenula fabianii J640. The promoter and terminator regions of the gene encoding glucoamylase from H. fabianii J640 were used for a new expression vector pHFGE-1. The performance of pHFGE-1 was compared with that of the widely used pG-1 transformant vector. H. fabianii J640 HF-TAMY cells were transformed with pHFGE-1 and Saccharomyces cerevisiae YPH-499 SC-TAMY cells were transformed with pG-1 both of which carried the Taka-amylase. Expression of Takaamylase by HF-TAMY showed higher than that by SC-TAMY. By using this new system we bred the new wastewater treatment yeast that shows a-amylase activity. This yeast appears to grow well under experimental wastewater conditions and is effective in treating model wastewater containing soluble and insoluble starch. Introduction Many food factories use wastewater treatment systems that use yeasts Yoshizawa 1978 1981 Sato et al. 1986 Moriya et al. 1990 Suzuki et al. 1991 Suzuki et al. 1996 . However some wastewater-containing polysaccharides such as raw starch and hemicellulose are difficult to treat because presently used yeasts secrete few enzymes that can digest these polysaccharides. One way to treat these wastewaters is to transform conventional wastewater treatment yeasts with the genes for polysac-charide-digesting enzymes such as raw starch-digesting a-amylase and acid xylanase. To this end we isolated Cryptococcus sp. S-2 Iefuji et al. 1994 which secretes several enzymes including raw starch-digesting a-amylase Iefuji et al. 1996a acid xylanase Iefuji et al. 1996b lipase Kamini et al. 2000 and polygalacturonase. We then obtained the genes that encode the raw starch-digesting a-amylase and .

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