The conventional alcohol fermentation method requires a large amount of energy for cooking the starchy raw materials prior to saccharification. The aim of this study was to compare the possibility of large scale alcohol fermentation from cassava slices were compared in low and high temperature cooking systems. The same amount of saccharifying and liquefying enzymes were used for cooking at low and high temperature. At low temperature cooking, conversion of glucose consumed in fermented mash to alcohol was 0.468g alcohol per g glucose of which was higher yield than that obtained at high temperature.

During the enzymatic production of L-phenylalanine exploiting L-phenylalanine ammonia lyase(E.C 4.3.1.5) and trans-cinnamic acid, the conversion yield of L-phenylalanine and the stability of L-phenylalanine ammonia-lyase per so or induced Rhodotorula glutinis IFO 0559 were investigated. And the glycerol added to the conversion reaction as stabilizer had effect only on L-phenylalanine and made it possible to obtain the 80％ conversion yield from trans-cinnamic acid. In addition, the more rapid and reliable method than the thin layer chromatography for determining the conversion yield will be disscused.

Poly(ethylene glycol)-PPB two phase system was used tot the purification of -galactosidase from Lactobacillus sporogenes. The smaller the molecular weight of concentration of PEG phase in-creased, proteins as well as -galactosidase was partitioned into the top phase. All cell debris were confined to the potassium phosphate phase (bottom phase), approached to the binodial line. The purification ratio increased by changing the polymer-salt composition of the tie line towards higher salt concentrations. It was also possible to obtain higher purification of the enzyme after two-step extraction using PEG 1000 and PEG 300. The top phase contained 74% of the total -galactosidase with a purification factor of 2.1.

The optimal conditions for the protoplast formation and regeneration of Streptomyces mitakaensis have been investigated. S. mitakaensis cells were converted to protoplast by treating with 0.1 mg/ of lysozyme in phosphate-tris buffer (pH 7.2) to the cells grown at the late logarithmic growth phase in the GBYN medium (gycerol 20g, beef extract 5g, yeast extract 5g, NaCl 5g in 1 liter of distilled water) contained 0.5% glycine. Cell regeneration from protoplast was accomplished in 10 days post inoculation on the R2 regeneration agar medium and at 3 days post inoculation on the H2 regeneration liquid medium. The efficiency of the regeneration was 0.l% in 3 days at 35.

The protoplast formation of Streptomyces mitakaensis was monitored with scanning electron microscopy and transmission electron microscopy. The normal cells formed regular mycelium and spore, and their cell wall and cell membrane appeared to be normal, but the cell wall of the lysozyme treated cells (1 mg/) was damaged, which was finally disappeared from cells to become protoplast in 30 to 60 minutes.

Two phase reaction system was used to hydrolyze the olive oil for fat splitting. Kinetics of lipases in two phase system were investigated by determining the hydrolysis rate of triglycerides at various olive oil concentrations in isooctane using the microbial lipases from Candida rugosa and Rhizopus arrhizus. The rate equation in lipid hydrolysis for various olive oil concentrations in two phase system was deviated from the Michaelis-Menten kinetics. The results suggested that the olive oil concentration in isooctane affects the interfacial area. The dependency of the interfacial area on olive oil concentration is greater at the lower olive oil concentration than at the higher substrate concentration. We modified the rate equation by considering the interfacial area between two phases depending on the olive oil concentration in solvent phase.

As a trial method of breeding L-lysine producing strains, the intraspecific protoplast fusion bet-ween Brevibacterium flavum ATCC 21528R and Brevibacterium flavum ATCC 21529S and the intergeneric protoplast fusion between Brevibacterium flavum ATCC 21528R and Corynebacterium glutamicum ATCC 13058S were performed. The optimum conditions for protoplast formation of these strains were examined and the effect of plasma expander on regeneration and/or fusion was also observed. Both fusants No. CH23 and No. CH4l showed higher productivity of L-lysine than those of parental cells under the optimum cultural conditions at a rate of 21％ and 8.9％, respectively. And, activity of several enzymes in L-lysine biosynthetic pathway including aspartokinase, a rate-limiting enzyme, was determined. Besides, metabolic control mechanism of L-lysine biosynthesis in fusant No. CH23 and in No. CH41 was investigated to compare with that of parental strains.

Optimal pH temperature and sucrose content for the production of vitamin B and riboflavin by Bacillus megaterium and Enterobacter aerogenes was studied by microbiological analysis. Optimal pH for the production of B was 6.0 by B. megaterium while the pH for E. aerogenes was 5.0. However, upon the addition of sucrose the optimal pH for B. megaterium shifted to 7.5 but E. aerogenes remained at pH 5.0. In the absence of sucrose, pH did not influence the yields of riboflavin produced by either bacterium. Addition of sucrose stimulated synthesis of riboflavin by both bacteria. Temperature had little effect on the production of vitamins by either bacterium.

In order to cloning of the nif genes of Enterobacter agglomerans NFB-264, the digested total DNA of the strain was ligated to pBR 322 and transformed into E. coli. Through the negative selection and colony hybridization, the transformants were obtained. The recombinant plasmids, pNEL 10 and pNES 20 were extracted from these transformants. It was known from Southern hybridization that pNEL 10 contained the 12 Mdal foreign DNA fragment hybridized with nif Q-X probe and pNES 20 included the 5 Mdal foreign DNA fragment hybridized with nif NE and nif YK probe.

Lipases from Candida rogosa and Rhizopus arrhizus were immobilized by entrapment with photo-crosslinkable resin prepolymer for the study of fat splitting and interesterification in isooctane-two phase system. Dioctylsulfosuccinate was selected as the most suitable surfactant during the immobilization. Lipase entrapped with hydrophobic photo-crosslinkable resin prepolymer(ENTP-3000) exhibited the highest activity, whereas lipase entrapped with hydrophilic gel(ENT-4000) was more stable in organic solvent. As the degree of hydrophobicity of the immobilization matrix was increased, Vm(app) of the lipase entrapped was increased, but Km(app) was approximately constant. While the optimum pH of the lipases entrapped on hydrophilic gel (ENT-4000) were around pH 7.0 for Candida lipase and Rhizopus lipase, the reaction rate of the lipases entrapped on hydrophobic gel were less dependent on pH variations for short reaction time. However, for longer reaction time, the lipnses from C. rugosa and R. arrhizus entrapped on hydrophobic gel yielded maximum rate at pH 6.0 and 6.5, respectively, Entrapment method endowed the lipase with thermal stability.

Aspergillus sp. (MK-24) producing a biological active substance that inhibited the venom proteinase activity was isolated from soil. The substance also inhibited the activity of trypsin and coagulation of blood, but did not inhibit papain, -chymotrypsin and pepsin. The substance was partially purified from culture filtrate by precipitaion with acetone, and by chromatography of DEAE-Sepadex A-50 column and Amberlite IRC-50 ion exchange. The inhibitory substance was stable in the wide pH range from 2.0 to 12.0 at 37, but not stable at in the alkaline pH. Only 12% of the activity was decreased by the heat treatment at 10 for two hours. The inhibition on venom proteinase (Agkistrodon bromohoffi brevicaudus) was a mixed type. The inhibitory activity depended on the preincubation time and completely depressed by cupric, zinc and cobalt ions. The inhibition on the venom proteinase was appeared strongly on casein but not on ovalbumin or hemoglobin as a substrate.

Aspergillus sp. MK-24 was cultured at 3 in the medium consisting of 2% glucose, 0.2% NaNO, 0.02% HPO, 0.02% MgSO 7, 0.02% KCl, and at initial pH of 5.0. The production of the inhibitor on venom proteinase reached to the maximum in 7 days. Sodium nitrate or potassium nitrate as a nitrogen source was favorable. The production of inhibitor was not affected by the addition of most of the inorganic salt used but depressed by lead, zinc, cobalt, mercuric or silver salts.