• 한국미생물·생명공학회지
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• 제25권1호
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• pp.68-74
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• 1997

The conversion of D-sorbitol to L-sorbose by Gluconobater suboxydans was analyzed, and continuous production of L-sorbose was carried out in immobilized cell reactors. L-Sorbose production by high densities of resting cells was more effective than by conventional batch fermentations. Sorbitol dehydrogenase, an enzyme converting D-sorbitol to L-sorbose, did not suffer from substrate inhibition, but from product inhibition. When L-sorbose production was carried out with Ca-alginate-immobilized cells, about 60 g/l of L-sorbose was obtained. On the other hand, when the corn steep liquor (CSL) concentration of medium was reduced to 0.08%, 80 g/l of L-sorbose was obtained. Outgrowth inside the immobilized carriers was thought to block the pores of the carriers so that substrate could not easily diffuse through the carriers. Continuous production of L-sorbose was well accomplished in a bubble column reactor, and 6. 5 g/l.h of productivity and 81.2% of yield were obtained at a substrate feeding rate of 0.08h$^{-1}$ under the optimum conditions with carrier volume of 55% and aeration rate of 3 vvm.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권5호
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• pp.340-344
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• 2000

Microbial oxidation of D-sorbitol to L-sorbose by Acetobacter suboxydans is of commercial importance since it is the only biochemical process in vitamin C synthesis. The main bottleneck in the batch oxidation of sorbitol to sorbose is that the process is severely inhibited by sorbitol. Suitable fed-batch fermentation designs can eliminate the inherent substrate inhibition and improve sorbose productivity. Fed-batch sorbose fermentations were conducted by using two nutrient feeding strategies. For fed-batch fermentation with pulse feeding, highly concentrated sorbitor (600g/L) along with other nutrients were fed intermittently in four pulses of 0.5 liter in response to the increased DO signal. The fed-batch fermentation was over in 24h with a sorbose productivity of 13.40g/L/h and a final sorbose concentration of 320.48g/L. On the other hand, in fed-batch fermentation with multiple feeds, two pulse feeds of 0.5 liter nutrient medium containing 600g/L sorbitol was followed by the addition of 1.5 liter nutrient medium containing 600g/L sorbitol at a constant feed rate of 0.36L/h till the full working capacity of the reactor. The fermentation was completed in 24h with an enhanced sorbose productivity of 15.09g/L/h and a sorbose concentration of 332.60g/L. The sorbose concentration and productivity obtained by multiple feeding of nutrients was found to be higher than that obtained by pulse feeding and was therefore a better strategy for fed-batch sorbose fermentation.

• KSBB Journal
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• 제8권1호
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• pp.10-16
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• 1993

The use of Jerusalem artichoke containing $\beta$-1, 2-fructose oligomer for the production of D-sorbitol and L-sorbose has been studied. The employment of inulinase(0.398%, v/v) for the hydrolysis of 40% (v/w) Jerusalem artichoke juice resulted in 36.7g/1 of glucose and 85.3g/1 of fructose at $50^{\circ}C$. These sugars were utilized as substrates for D-sorbitol and L-sorbose production. Coimmobilization of inulinase and permeabilized cells of Zymomonas mobilis in the mixture of chitin (5%, w/e) and x-carrageenan(4%, w/v) resulted in the production of 30.2g/1 of D-sorbitol by using inulin as a substrate. The process of L-sorbose production from D-sorbitol by Gluconobacter suboxydans was optimized with respect to the substrate concentration, level of dissolved oxygen and glucosic and concentration. Gluconlc acid produced by Zymomonas mobilis from glucose was found to inhibit Gluconobacter suboxtans in conversion of D-sorbitol to L-sorbose. In view of removing such inhibitory effect by gluconic acid, mutants were selected by the NTG (N-methyl-N'-N'-nitro-N-nitrosoguanidlne) treated method. Mutants selected by NTG mutagenesis showed no inhibitory effects of gluconic acrid against L-sorbone production when its concentration increased up to 100g/1. A mutant produced 40.1g/l of L-sorbose in the medium containing 100g/l D-sorbitol and 100g/l-gluconic acid. This result is consider able when compared with L-sorbose concentration (21.7g/1) obtained from the fermentation with wild type strain of Gluconobacter suboxnians.

• Applied Biological Chemistry
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• 제36권6호
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• pp.481-487
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• 1993

Acetobacter suboxydans의 휴지균체를 이용한 sorbitol로부터 sorbose 생산계에 대해 검토한 결과 5% sorbitol농도에서 균체를 약 6 mg/ml 농도까지 첨가할 때 sorbose의 생산량이 급격히 증가하였다. Sorbose 생성의 최적 반응 온도는 $30^{\circ}C$, 최적 pH는 6.0이었으며 금속 이온은 1 mM $Al^{+3}$ 이온에 의해 약 12%의 증가를 보인 반면에 $Ni^{+2}$ 이온 첨가시 현저한 sorbose 생산 저해를 보였다. p-aminobenzoic acid를 1.0 mM 첨가시 약 20%의 sorbose생성이 증가되었으며, Ca-pantothenate 첨가는 감소 효과를 나타내었으나 p-aminobenzoic acid와 혼합첨가에 의한 sorbose 생성은 약 7% 증가하였다. 총 반응액 1.5 ml를 50 ml 삼각 프라스크에서 반응시켰을 때, 기질 5% sorbitol은 20시간만에 완전히 sorbose로 전환되었다.

• Journal of Microbiology and Biotechnology
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• 제8권6호
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• pp.696-699
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• 1998

Biological oxidation of D-sorbitol to L-sorbose using permeated and immobilized cells of Gluconobacter suboxydans was carried out to investigate the optimum reaction condition. The stabilization effect of cross-linking agents such as glutaraldehyde, tannic acid, and polyethylene imine to prevent the leakage of enzymes from beads containing permeated and immobilized cells of G. suboxydans was examined by the production of L-sorbose from the mixture of D-sorbitol and gluconic acid. The protein concentration effused from immobilized beads treated with only glutaraldehyde was $5.2\mug/m\ell$ after 20 h. The beads of G. suboxydans immobilized with alginate and cross-linked with 0.3% glutaraldehyde was the most useful for the oxidation of D-sorbitol to L-sorbose.

• KSBB Journal
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• 제5권1호
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• pp.1-8
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• 1990

D-sorbitol을 첨가한 배지에서 G. melanogenus를 배양하였을 때에 D-sorbitold은 L-sorbose를 거쳐 2 keto-L-gulonic acid로 전환되었다. G. cerinus를 D-sorbitol 대신에 포도당을 첨가하여 준 배지에서 배양하였을 때에 포도당은 2-keto-D-gluconic acid, 5-keto-D-gluconic acid, 2.5 diketo-D-gluconic acid로 산화되었다. 2,5-diketo-D-gluconic acid는 포도당이 2-keto-D-gluconic acid를 거쳐 생성되는 것으로 확인되었다. 배양액에 첨가하여 준 탄산칼슘의 양을 증가시키면 5-keto-D-gluconate의 생성이 증가하였고, 탄산칼슘대신에 다른염기를 배양액에 가하여 주었을 때는 2keto-D-gliconate만 생성되었다.

• 약학회지
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• 제32권6호
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• pp.386-393
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• 1988

Chemical transformation of D-glucose to 2-keto-L-gulonic acid and L-ascorbic acid has been examined. D-Sorbitol obtained from D-glucose was microbiologically oxidized to L-sorbose by G. suboxydans in 90% yield. On treatment of L-sorbose with acetone in the presence of sulfuric acid, its diacetonide is obtained in 95% yield. This diacetonide is oxidized to the corresponding acid with nickel chloride-hypochlorite, and the acid is directly transformed to L-ascorbic acid. The over all yield of Vitamin C from D-glucose achieved is 54%.

• KSBB Journal
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• 제7권1호
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• pp.15-20
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• 1992

식품첨가물 등으로 이용 되어지는 sorbitol을 $\beta$-1,2fructose oligomer를 함유하고 있는 Jerusalem artichoke를 이용하여 생산 하였으며, 이는 L-sorbose생산에 있어서 전구물질로 이용된다. Inulinase와 oxidoreductase의 동시고정화여, inulin을 와 fructose로 가수분해함과 동시에 가수분해산물을 sorbitol로 전환시켰다. Inulinase와 oxidoreductase는 chitin(5%, w/v)과 K-carrageenan(4%, w/v)으로 고정화 시켰다. 0.2% CTAB(Cetyltrimethylammonlumbromide)처리에 의해서 투과성이 향상된 Zymomonas mobilis세포내의 함유 되어있는 oxidoreductase의 activity가 향상되었다. Jerusalem artichoke juice를 acid에 의한 가수분해에 의해서 40%(v/w) juice 100ml당 53.64의 total carbohydrate yield를 얻었고, 온도 변화에 따른 가수분해시 $85^{\circ}C$에서 반응이 신속히 일어났다. Inulinase를 이용한 가수분해에 의하여 35.56g/l의 glucose와 85.32g/l의 fructose가 생성되었으며, sorbitol생성에 기질로 이용되었다. Inulinase와 Z. mobilis세포의 동시고정화 반응기에서 35.15g/l의 sorbitol을 생성하였으며, 이는 동시고정화를 하지 않았을때의 sorbitol생성농도(35.64g/l)보다 낮았다.

• 식품산업과 영양
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• 제2권1호
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• pp.16-21
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• 1997

Microbial of enzymatical methods are suitable for production of rare monosaccharides. Using oxidation and reduction ability of Microorganisms, various rare ketoses and polyols can be produced, for example D-tagatose from galagtitol by Enterobacter agglomerans strain 221e. L-tagatose from galactitol by Klebsiella pheumonias strain 40b, L-psicose from allitol by Gluconobacter frateurii IFO 3254, D-talitol from d-tagatose by Aureobasidium pullulans strain 113B, allitol from D-psicose by Enterobacter agglomerans strain 221e and so on. We can produce various rare aldoses and ketoses using aldose isomerases, for example L-galactose from L-tagatose by D-arabnose isomerase, and L-ribose from L-ribulose by L-isomerase, and so on. D-Tagatose 3-epimerase of Pseudomonas sp. ST-24 is very useful for preparationof various rare ketoses, for example D-psicose from D-fructose, D-sorbose from D-tagatose, L-fructose, from L-psicose and so on. Using polyol dehydrogenases, aldose isomerases and D-tagatose 3-epimerase, we can design the suitable for production of a certain rare monosaccharide from a suitable substrate.

• 한국환경농학회지
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• 제32권3호
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• pp.240-243
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• 2013

효모의 유용 기능을 탐색하기 위해서는 효모만을 스크리닝 하는 방법의 정립이 요구되는데 이를 위해서 알로에에 정착하는 효모 군집을 분석하였다. 본 연구에서는 알로에 베라(Aloe vera)와 알로에 사포나리아 (Aloe saponaria)의 잎을 파쇄하여 세균을 억제하기 위해서 항생제 chloramphenicol과 streptomycin과 곰팡이 생장을 억제하기 위해서 Triton X-100과 L-sorbose를 포함한 4가지 선택 배지(DOB with CS, GPY, YM, SCG)에 도말 하였고, A. vera에서 총 67균주, A. saponaria에서 총 42균주를 분리해 내었다. 분리된 균주를 ITS 1, 4 primer를 사용하여 sequence의 계통분석을 실시한 결과, A. vera에서는 Meyerozyma 가 56균주, Rhodotorula 가 1균주, Sporobolomyces 가 1균주, Cryptococcus 가 9균주, A. saponaria에서는 Rhodosporidium 가 41균주, Sporobolomyces가 1균주로 분포 되었다. 계통분석한 모든 분리 균주가 효모로 밝혀졌으며, 특히 대형 플레이트를 도입하여 신속하고 간편하게 대량의 효모를 발굴할 수 있음을 보였다.