Characterization of Brewing Yeast Expressing Glucoamylase Selected by Rare Mating.

Rare Mating에 의한 양초효모에서의 glucoamylase 발현 균주 HCS 선별 및 특성

Rare mating was used to select a respiratory deficient mutant of Saccharomyces cerevisiae HDC52 strain. Glucoamylase gene of S. diastaticus K114 was developed into the RD mutant which could uptake maximum amount of non-fermentable sugars through the expression of glu- coamplyase gene and the fermentation characteristics of the developed strain HCS were investigated. The size of HCS yeast and HBD52 yeast strain were 13 $\mu\textrm{m}$ and 10$\mu\textrm{m}$ respectively. HCS strain which can uptake maximum amount of non-fermentable sugar through the expression of glucoamylase gene was developed. By karyotype anal- ysis. HCS stain but not RD mutant HBC52 showed a band of 1150 kb chromosome DNA This band should include glcoamylase gene from Saccharomyces diataticus K114 THis strain has glucoamylase which can degrade starch By transduction and contrnuance of glucoamylase gene HCS strain gegraded strach and formed halo. Also, HCS strain maintained the character after 50 generations. Glucoamylase activities of Saccharomyces diastaticus K114 and HCS yeast strains are 9.5 and 2.7~3.4(unit/ml) HCS and HBC52 strain showed similar sugar fermentation patterns and low flocculation In spore and film forming test, HCS and HBC52 strain formed neither spores nor films. In the limit fermentation test, HBC52 strain showed fermentation level of 68% and HCS strain showed 76~78% As the limit attenuation of HBC52 and HCS were ($2.00^{\circ}$P) and ($0.7~0.93^{\circ}$P) This study demon- strates and HCS strain may be used for low carbohydrate beer fermentation.

S. cerevisiae HBC52와 S, diataticus K114 의 rare mating 에 의해 개발된 HCS 균주들은 크기가 약 $13\mu\textrm{m}$ karyotype 분석결과 K114 균주에만 있는 약 1150kb 분자량을 가지는 염색체 band를 유지하였으며 전분을 분해하여 halo 를 형성하였다. Glucoamylase 활성은 약 2.7~3.4 unti/ml 를 가진 균주임이 밝혀졌으며 당 발효실험과 응집성 실험을 수행한 결과 HBC52 균주와 유사한 당 발효특성을 보이고 응집성 특성도 약응집성의 floculation type으로 비슷하였다. 그리고 HCS 균주의 포자형성과 피막형성 유무 실험에서는 양조효모인 HBC52 균주와 같이 포자가 형성되지 않았으며, 피막도 형성되지않았다. 균주들의 최종당도 실험은 HBC52균주가 약 68%의 발효수준을 나타냈고, HCS 균주들은 이 보다 높은 76~78%의 수준을 보였따. 즉 HBC52 균주가 최종당도($ 2.00^{\circ}$P)를 보인 반면 HCS 균주들은 ($0.7~0.93^{\circ}$P)를 보이는 결괄르 나타내어 맥주양조에서 low carbohydrate beer를 생산할 수있음이 확인되었다.