각종 시료(各種 試料)로부터 구연산생성능(酸生成能)이 있는 효모(酵母)를 검색(檢索)하여 우수균주(優秀菌株)로서 Strain No. 13을 선정(選定)하고 이를 동정(同定)함과 아울러 구연산(酸) 생산조건(生産條件)을 검토(檢討)한 결과(結果)는 다음과 같다. 1) 선정균주(選定菌株)는 Lodder의 분류법(分類法)에 의(依)하여 Hansenula anomala var. anomala로 동정(同定)되었다. 2) 10% glucose를 함유(含有)하는 기본배지(基本培地)에 있어서 구연산(酸) 생산(生産)의 최적조건(最適條件)은 $CaCO_3$ 첨가농도(添加濃度) 3%, 배양온도(培養溫度) $30^{circ}C$, 진탕속도 110 oscillus/min 이였다. 3) 배지(培地) 중(中)의 질소원(窒素源)으로서는 0.1% $NH_4Cl$이 가장 좋았으며 peptone 등(等)의 유기질소원(有機窒素源)은 균주생육(菌株生育)에 있어서 $NH_4Cl$보다 좋았으나 구연산생산(酸生産)은 저하(低下)되었다. 4) 당질(糖質)로서 glucose를 사용(使用)하는 경우 구연산(酸)의 대당수율(對糖收率)은 glucose 농도(濃度) 10% 정도(程度)에서 가장 양호(良好)하였다. 5) 채종유박(菜種油粕), 들깨박(粕) 탈지미당(脫脂米糖) 등(等)은 구연산생성(酸生成)을 증가(增加) 시켰으며 0.2% yeast extract 사용시(使用時)에 비(比)하여 5% 채종유박(菜種油粕) 추출액(抽出液) 사용시(使用時)는 구연산(酸)이 약(約) 40% 증산(增産)되었다. 6) 배지(培地) 중(中)의 $KH_2PO_4$의 최적농도(最適濃度)는 0.05%, $MgSO_4{cdot}7H_2O$의 최적농도(最適濃度) 0.025% 이였다. 7) 최적조건하(最適條件下)에서 균(菌)의 생육(生育)은 5일(日)까지, 구연산생성(酸生成)은 6일(日)까지 계속(繼續) 증가(增加)하였으며 6일(日) 배양시(培養時) 대당(對糖) 46%의 구연산(酸)을 얻었다.
A potent citric acid producing strain was selected by an extensive screening test of the yeasts isolated from the various sources. These experiments were conducted to identify the selected strain and investigate the cultural conditions for citric acid production. The results obtained were as fellows: 1. The selected strain of yeast was identified to Hansenula anomala var. anomala by a taxnoomic study of Lodder. 2. The optimum conditions for citric acid production in the basal medium containing 10% glucose were: temperature $30^{circ}C$, the concentration of $CaCO_3$ 3% and the velocity of oscillation 110 oscills/min. 3. As a nitrogen source ol the basal medium $NH_4Cl(0.1%)$ was the most effective for citric acid production. Organic nitrogen sources such as peptone were adequate for growth of the strain but not for citric acid production. 4. The most effective concentration of glucose was 10% in yield ratio of citric acid from sugar. 5. The addition of defatted rape seed, defatted perilla or defatted rice bran to the medium was effective for citric acid production. When 5% extract solution of defatted rape seed was added, the citric acid production was increased as much as 40% as compared with the case of adding yeast extract(0.2%). 6. The most effective concentration of $KH_2PO_4$ and $MgSO_4{cdot}7H_2O$ in the medium(for citric acid Production) was 0.05% and 0.025% respectively. 7. Under the optimum cultural conditions, the growth of the strain was continued up to 5 days and the increase of citric acid production was continued up to 6 days, showing the yield ratio of 46% to glucose.
