Production of Rhizobium meliloti M14 Inoculum by Semi-continuous Cultivation

반연속식(半連續式) 배양(培養)에 의(依)한 Rhizobium meliloti M14의 균체생산(菌體生産)

As a basic studies for the laboratory scale production of alfalfa inoculum, Rhizobium meliloti M14 was characterized for its carbon and nitrogen sources, and some parameters for broth cultivation in a chemostat were studied by semi-continuous operation. The result s obtained were as follows. 1. Growth rate of the strain was increased by disaccharides than by monosaccharides tested, and pentoses resulted in poor growth than hexoses. Sugar alcohols including inositol supported the best growth among sugars. 2. Mannitol in the yeast-mannitol-broth was substituted by natural carbon sources such as malt extract or molasses. 3. Ten per cent of fresh yeast water appeared to supply enough amount of growth factor s for the strain, and the effect was equivalent to 0.24 percent of the commercial yeast extract powder. 4. Batch growth of the stain in a chemostat, New Brunswick Micro Ferm 28L, reached in the early stationary growth phase of $5{\sim}7{\times}10^9cells/ml$ after 36 hours of incubation. The culture at this stage was switched to semi-continuous cultivation, and the culture broth of four-fifth of the working volume was recovered every 24 hours when the maximal count was obtained.

전보(前報)에서 분리(分離)한 Rhizobium meliloti M14를 공시(供試)하여 그 종균생산(種菌生産)을 위한 기초적(基礎的) 연구(硏究)로서, 탄소원(炭素源) 및 질소원(窒素源), chemostat의 반련속식(半連續式) 운용(運用)에 의한 액체배양(液體培養)을 시도(試圖)하여 아래와 같은 결과(結果)를 얻었다. 1. 탄소원(炭素源) 비교(比較)한 단당류(單糖類), 이당류(二糖類) 및 당(糖)알콜류(類) 가운데 당(糖)알콜류(類)에서 가장 생육(生育)이 높았고 단당류중(單糖類中) 오탄당(五炭糖)에서의 생육(生育)은 육탄당(六炭糖)보다 저조(低調)하였다. 2. 천연(天然) 탄소원중(炭素源中) malt extract 및 molasses에서의 생육(生育)은 mannitol에서의 생육수준(生育水準) 이상(以上)이었다. 3. 질소원(窒素源)으로서 yeast water에 대(對)한 편향성(偏向性)이 인정(認定)되었으며 그 10%에서의 생육수준(生育水準)은 yeast extract 0.24%와 상응(相應)하였다. 4. Chemostat (MicroFerm 28L, New Brunswick model CMF-128S) 의 단식배양(單式培養)에 의(依)하여 36시간(時間) 배양(培養)으로 생균수(生菌數) $5{\sim}7{\times}10^9cells/ml$ 수준(水推)의 stationary phase에 도달(到達)하였고, 반련속식(半連續式) 운용(運用)으로 매(每) 24시간(時間)마다 동일수준(同一水準)의 생균(生菌)을 가진 배양액(培養液)을 그 운용량(運用量)에 대(對)하여 5분(分)의 4를 회수(回收)할 수 있었다.