Aspergillus niger KCTC 6144 포자를 alginate bead로 고정화하여 호박배지(포도당 9%, 호박가루 1%, pH 6)를 이용하여 구연산 발효를 수행하였다. 고정화된 A. niger 포자가 bead 내에서 발아하여 균사가 bead를 뚫고 자라났으며 3$0^{circ}C$에서 4일간 배양 후 그 크기는 2.0~2.5mm에서 6~8mm로 커졌다. 3$0^{circ}C$에서 5일간 진탕배양(150rpm)으로, 50ml 호박 배지가 든 250ml 삼각 flask를 사용하여, 구연산 생성의 최적 발효 조건을 검토한 결과는 다음과 같다. 탄소원으로 포도당을 12%로 올렸을 때 구연산의 생성이 최고에 달하였고, 질소 및 무기질원으로 1%의 호박가루가 구연산 생성에 가장 알맞는 농도였다. 초기 pH는 6.0, bead의 수가 100개로 하였을 때 구연산 생성이 가장 좋았으며, 포도당 12%, 호박가루 1%로 한 최적 배양 조건에서, 5일만에 구연산이 23.5g/ι로 최고로 생성되었다.
The spore of Aspergillus niger KCTC-6144 was immobilized on alginate gel beads. When pumpkin powder was used with glucose for a medium of citric acid fermentation by Aspergillus niger beads, the beaded Aspergilus niger grew up inside the bead and mycelia penetrated through the pore of the bead membrane. The bead size became largely from 2.0∼2.5mm to 6∼8mm after growing at 30$^{C}$ for 4 days. Studies of optimum culture conditions on citric acid fermentation using Aspergillus niger beads on pumpkin medium (pumpkin powder 1% +glucose 7%, pH 6.0) were carried out in submerged cultures on 250m1 Erlenmeyer flask. As a result, it was found that to reinforce 12% as carbon source was good for citric acid production and that 1% pumpkin powder was good as nitrogen and mineral source in orbital shaker (150rpm) at 30$^{C}$ for 5 days. The optimum initial pH on citric acid production was pH 6.0 and it was found that 100 beads of immobilized Aspergillus niger was adequate for citric acid production in a 250ml Erlenmeyer flask containing 50m3 of pumpkin medium solution with orbital shaker at 30$^{C}$ for 5 days. We also found that maximal production of citric acid was 23.5g/ℓ at optimal condition (at 30$^{C}$ for 5 days, pH 6.0, and 100 beads and medium containing 1% pumpkin powder plus 12% glucose).
