We have studied the relation between Ca$^{2+}$-ATPase and trigger peptidase(TPase) which are membeane protein well known as their significant role for signal transduction of mating pheromone in heterobasidiomycetous yeast. Rhodosporidium toruloides. We found out that there were Ca $^{2+}$-ATPase and TPase together in isolated calmodulim binding protein(CBP), usion calmodulin affinity column chromatography after solubilization of mation type a cell membrane protein, and that the dependence of enzyme activity of both the enzymes on Ca$^{2+}$, phospholipid and nonionic detergent are similar. However, Ca$^{2+}$-ATPase hed quite absolute dependence on calmodulin and, on the other hand, TPase didn't have any dependence. Judging from the fact that there are both enzymes in CBP which the dependence of calmodulin are quite different, we found out that both enzymes were made to their compound and existed in mating type a cell membrane.

One bacterial strain, that had made the largest inhibition zone at the antagonism assay and also that lost the inhibition activity by the protease treatment, was isolated from raw milk. That strain was identified as Streptococcus salivarius subsp. thermophilus. The specific growht rate of this strain was maximum at 45$\circ $C. However, at this temperature the strain produced no bacteriocin. The bacteriocin activity was quite stable even at high temperature. Moreover, the activity of the vacteriocin was sensitive to proteases. but not to $\alpha $-amylase, DNase I, or RNase.

We estabilished the procedure for isolation of Streptococcus salivarius subsp. thermophilus from raw milk. First, urease-producing lactic acid bacteria in raw milk were screened on the HY agar medium containing urea. Thereafter the urease-producing colonies were tested the ability to ferment maltose and to grow at43$\circ $C. We obtained about 400 maltose-negative colonies that grew at 43$\circ $C. No significant difference in carbohydrate fermentation test for isolated and type strains(S. salivarius subsp. thermophilus ATCC 19258 and ST-4) was found. And all of the isolated strains were able to ferment galactose. Furthermore, it was investigated that the cellular fatty acid profiles of isolated strains were similar to that of type strains. These results indicated that the isolated strains from raw milk were S. salivarius subsp. thermophilus. But when the isolated and type strains were incubated in 12% reconstituted skim milk at 43$\circ $C, the isolates produced lactic acid more slowly than the type strains.

Enterococcus faecalis KBL703 has three plasmids(p703/9, p703/5 and p703/4). Within p703/5, the specific DNA region that would confer replication function(replication origin) was searched by transformation experiments. In order to use as the recipient of transformation, two plasmid-cured strainsd were made from this strain. Four recombinant DNA constructs, each containing fragment of p703/5 and CAT(chloramphenicol acetyl transferase) gene were also made. And they were used to transform the plasmid-cured strains. Only one DNA construct containing 3.6 kb SalI fragment was stably maintained as plasmid in these strains. Additional experiment using another Enterococcus faecalis strain(ATCC29212) as a recipient was successfully done and it was confirmed that this newly constructed recombinant plasmid plasimid contained the replication origin from p703/5 plamid.

Intergeneric hybrids formed between Aspergillus oryzae var oryzae and Penicillium verruculosum F-3 were obtained by nuclear transfer technique. Several auxotrophic mutants isolated fromconidio-spores of the two strains mutagenized with ultraviolet and N-methyl-N-nitrosoguanidine. Optimal conditions for formation of intergeneric hybrids were investigated. Frequencies of hybrid formation by nuclear transfer were 3$\times $10$^{-5}$ $~$1$\times $10$^{-5}$. From observation of genetic stability, conidial size, DNA content, nuclear stain, it was suggested that their karyptypes are aneuploid. The hybrid posses the 1.3$~$2.2 fold higher amylase activities than those of parental strains. It was also revealed that some hybids had different isozyme patterns compared to those of parental strains by amylase activity assays.

From the pSKH201 plasmid which had been previously cloned in our laboratory, a 3.0kbp insert DNA encoding the alkaline phosphatase of Kluyveromyces fragilis was cleaved with several restriction endonucleases and ligated int the appropriate sites of M13mp18/19 vectors and sequenced by Sanger's dideoxy chain termination method. The sequence contained a 1,638 bp open reading frame(ORP) whose similarities in nucleotide, when compared with those of Saccharomyces cerevisiae and Escherichia coli by GENETYX program, were found to be 61% and 46%, respectively. The deduced amino acid sequence consists of 546 amino acids and contains several homologous regions in the alkaline phosphatases of E. coli, S.cerevisiae and human placenta.

The homology among the genes coding for degradation of bipheny(BP) and 4-chlorobiphenyl(4CB) was comparatively analyzed by Southern hybridization in several BP/4CB degrading bacterial strains. As the hybridization results of their genomic DNAs with pcbABCD as the DNA probe, the group of Pseudomonas sp. DJ-12. P08 and P27 strain was separated by the group of P20 and P1242 strains. The P. pseudoalcaligenes KF707 showed the hybidization signal which was homologous to the group of DJ-12, but they had different restriction endonuclease sites. The pcbAB genes in pCUl recombinant plasmid from Pseudomonas sp. DJ-12 appeared to be homologous to pchAB genes in pKTF20 cloned from P. pseudoalcaligenes KF707, but the C genes in both strains were not homologous. The bphABC in pKTF20 showed the signals homologous to the cbp ACB in pAW6194 cloned from P. putida OU83, but homologous signal was not found botween the pcbABCD genes in pCUl and the cbpADCB genes in pAW6194 recombbinant plasmid.

Antitumor polysaccharides were known to activate complement system and to increase specific serum proteins in mouse, and researcher reported that antitumor activity of polyasccharides might be correlated with their biological properties such as activation of complement system and increase of specific protein $L_{A}$, $L_{B}$ and $L_{C}$ within the mouse serum. In case of several Ganoderma lucidum, there was no correlation between their antitumor activities and their bioloical properties, but the antitumor activities against sarcoma 180 of the alkali extracted crude polysaccharide fractions of the Ganoderma lucidum IY 009. AS, T, AI and M were correlated with their bioloical properties such as anticomplementary activity and intensity of mouse serum protein $L_{A}$, $L_{B}$ and $L_{C}$.

The induced antibiotic peptides were isolatde from the white-spotted folwer chafer, Protaetia brevitarsis by injection of E. coli suspension to the larvae of the insect. The antibacterial activity of the peptides were assayed by the plate growth ingibition method, and were purified by ion-exchange chromatography, reversed-phase HPLC, ion-exchange HPLC and SDS-PAGE etc. The peptides were estimated as 9 kDa in molecular weight and named Protaecin I and Protaecin II, respectively. Protaecin I and II have strong antibacterial activities against Gram-positivie and/or Gram-negative bacteria, and they are stable in the heat treatment and in the range of pH 2-12.

The production and the characteristics of protease produced by Bacillus sp. SH-8 and Bacillus sp. SH-8M were investigated under the different pH conditions. Bacillus sp. SH-8 and Bacillus sp. SH-8M showed the maximum activity of protease at 60 hours(70 units/ml) AND 96 houre(50 units/ml) cultivation, respectively, under the alkaline condition(pH 10.2). However, Bacillus sp. SH-8M exhibited the maximum activities in 8 days cultivation at pH 6.9 and in 6 days cultivation at pH 7.7 Bacillus sp. SH-8M showed the protease activity at the pH change from alkaline to neutral condition, whereas Bacillus sp. SH-8 did not. In addition. all the enzymatic characteristics of protease produced by Bacillus sp. SH-8 and Bacillus sp. SH-8M were similar with the regardless of different pH conditions.

Purification and characterization of pectate lyase from Bacillus sp HSA-925. Bacillus sp. HSA-925 isolated from soil produced constitutively an extracellular pectate lyase when cultivated in LB broth. The pectate lyase(EC 4.2.2.2) was purified from the cuylture broth by preciptation with ammonium sulfate, followed by column chromatography on CM-cellulose C-50 and repeated gel filtration on Sephadex G-75G. The enzyme had a molecular weight of 32-33 kDa. The activity was mazimum at pH 9.5 AND 45$\CIRC $C. The enzume activity was stable at 55$\circ $C for 15 min and between pH7-12. The activation energy, Km and V$_{max}$ for the pectate lyase were 5.8779 kcal/mol, 6.33$\times $10$^{-2}$ mol/ml and 2.09$\times $10$^{2}$ $\mu $mol/min respectively. The enzyme was activated by Ca$^{2+}$, Cu$^{2+}$ and inhibited by Li$^{+}$, Hg$^{2+}$, EDTA.

The optimum cultural temperature and time for the pullulanase production by Bacillus cereus subsp. mycoides were 35$\circ $C and 48 hrs, respectively. The addition of egg albumin and casein to the basal medium increased the enzyme production. The enzyme was purified by ammonium sulfate fractionation and DEAE-cellulose column chromatography. specific activity of the purified enzyme was 82.37 U/mg protein and yield of theenzyume activity was 62.1%. The purified enzuyme showed a single band on ployacrylamide disc gel electrophoresis and its molecular weight was estimated to be 66.,000 by SDS-polyacrylamide disc gel electrophoresis. The isoelcular point for the purified enzyme was pH 5.0. The optimum temperature and pH were 50$\circ $C and pH 6.5, respectively. The purified enzyme was stable below 40$\circ $C and in the pH range of 6.5~10.0 The pullulanase activity was greatly inhited by Ag$^{+}$, Hg$^{2+}$ and EDTA, and its heat stability was increased by the addition of Ca$^{2+}$. The tydrolysis product with the enzyme on pullulan was maltotriose.

In order to induce the rapid alcohol fermentation through the increases of the cell density in a continuous alcohol fermentation of naked barley, the single-cultivation with S. cerevisiae IS-019(SCM, ordinary control), mixed-cultivation with Saccharomyces uvarum IS-026 having a flocculent ability and S. cerevisiae IS-019(MCM), and mash recirculation by single-cultivation of S. cerevisiae IS-019(MRM) modes were investigated. The cell mass in the mixed-cultivation mode was about 10% higher than that of ordinary control but the final alcohol yield was slightlyl decreased. When recycled the mash with the flow rate of 7 l/h from V$_{6}$ to V$_{5}$ fermentors under the ordinary control, the cell density was distributed at 140~170$\times $10$^{6}$ cell/ml depending upon the fermentorsorders, higher about 20% than that of the ordinary control. Under these conditions the alcohol productivity of the maximum and the overall was 12.16 g/l$\cdot $h with an alcohol of 7.6% at the V$_{5}$ fermentor and 1.19 g/l$\cdot $h with an alcohol of 8.94%, respectively. For higher cell mass it was more effective to apply the mash recirculation mode with the single-cultivation of S. cerevisiae IS-019 in a pilot scale multi-stage CSTR.

The characteristcs of monascus fermentation using a hyperpigment-producing mutant, Monascus sp. J101, were analyzed, and the extractive fermentations employing permeabilizing agents and resin were carried out to increase the productivity of red pigment. And the kinetic analysis was also carried out in case of the monascus fermentation using Amberlite XAD-7. The extracellular content of the red pigment produced by Monascus sp. J101 was about 17% of the total, and the production of pigment was regulated by its own product. The cell growth reached a stationary phase at 48 hours ofter inoculation, whereas the pigment production continued up to 100 hours, which showed the pattern of a mixed growth-associated type. During the fermentation, various permeabilizing agents were added to the culture medium and their effects on pigment production were examined. By adding 0.05% Triton X-100 at 48 hours of cultivation, about an 18% increase in pigment production was accomplished as compared to the control, 12% ethyle acetate and 15% for 0.05% deoxycholate, respectively. When a nonionic adsorbent, Amberlite XAD-7 was added to the culture medium at a concentration of 12.0% at 48 hours of cultivation, the pigment production was enhanced by about 48.9% as compared to the control.

The bacteria which secrete surface-active agent and decrease the surface tension of culture broth were isolated from soil samples. Among them, biosurfactant producing strain KK-7 was selected and emulsification was also detected. The KK-7 produced biosurfactant not only lipid but also glucose by using carbon source. Taxonomical characterization tests have demostrated the strain KK-7 to be Pseudomonas aeruginosa. The media composition of the P. aeruginosa KK-7 for the biosurfactant production was 1% glucose, 0.5% tryptone, 0.2% yeast extract, 0.15% potas sium phosphate mono-dibasic, 0.05% MgSO$_{4}$, initial pH 8.5, at 30$\circ $C for 2 days. In this condition, the concentration of biosurfactant was reached CMC 5 in the culture broth. Surface active material was produced maximum at stationary8 phase, but emulsification power was higher at log phase than stationary phase. It was considered that P. aeruginosa KK-7 produced biosurfactant more than one type having defferent properties and each maximum production time was different. The minimun surface tension of biosurfactant in 50 mM Tris buffer (pH8.0) was 28 dyn/cm, and CMC was 1 g/L.

In this study we compared traditional cheesemaking process with the process utilizing ultrafiltration(UF) system. The whole milk retentates were prepared by ultrafiltration to volume concentration ratio(VCR) of 2.00:1, 2.25:1 and 2.50:1. Along with the untreated whole milk, there were studies in terms of the change of pH, titratable acidity and Soxhlet-Henkel($\circ $SH) value by mesophilic lactic starter and curd formation by rennet during Gouda cheese manufacture. Due to the increase of buffering effect titratable acidity and $\circ $SH value increased with the higher concentration ratio. When inoculated with the same volume of mesophilic lactic starter, less pH change occurred in UF retentates than in control milk. When added 0.0025% rennet, UF retentates coagulated 16~ 17 minutes ealier then the control milk. Gouda cheese yield from raw milk and UF retentates was 12.5~13.1% equally, but yield efficiency of UF retentate cheese was slightly higher than that of the raw milk cheese. Quantity of whey from retentate cheese was inversely related to VCR. But whey from retentate cheese contained higher percentage of amjor components than that from control milk cheese. In early ripening, the concentrations of lactose and soluble nitrogen compound were higher in retentate cheeses. Lactose content of control milk cheese was 3.49% and that of 2.00:1. 2.25:1, 2.50:1 VCR retentate was 3.77%, 4.89%, 7.03%, respectively. Thus, the more concentrated cheese contained a higher amount of lactose and all the lactose was hyerolyzed durion 35-day ripenion period. Soluble nitrogen compound of control milk cheese was 1.22% and that of UF cheeses was 1.82~2.06%. After 20-day ripening, soluble nitrogen compound increased starply in UF cheese.

A novel method for production of concentrated purity maltose using swollen extruded corn starch was investigated. Degree of gelatinization of extruded starch suitable for maltose formation was found to be around 70%. The optimal amiunt of enzyme was 400 unit fungal $\alpha $-amylase per g of starch, and the reaction time was 12 hours. At extruded starch concentration of 300 g/l(w/v), maltose concentration and content were reached up to 220 g/l(w/v) and 77%(w/w), respectively. The maltose forming reaction was also successfully proceeded at high starch concentration of 700 g/l(w/v), however, the conversion yield and content were decreased. By the addition of extruded starch by fed-batch wise, the maltose concentration, purity, and conversion yield could be improved up to 465 g/l(w/v), 70%(w/w), and 0.63, respectively. The investigated maltose production process seems to have many potential advantages over the conventional process utilizing liquefied starch, and the feasibility for industrial application needs to be evaluated.