Kim, Jae-Young;Chung, Jin-Woo;Cho, Kyung-Yun;Yi, Yong-Sub
Microbiology and Biotechnology Letters
/
v.37
no.3
/
pp.183-188
/
2009
Seven isolates showing strong proteolytic activity, KYC 1028, 1100, 1134, 1139, 1151, 1159, and 1182, were collected. Out of them, the broth of KYC 1134 and KYC 1139 showed the high proteolytic activity measured by azocazein. To determine 16S rDNA sequences for identification, 16S rDNA of seven isolates were amplified and compared with the 16S rDNA sequences of other myxobacteria at NCBI. It is evident from the phylo-genetic tree that the isolates belong to the genus Myxococcus. Sharing high percentage similarity values with myxobacteria, the 16S rDNA sequences were involved in two species, Myxococcus macrospores and M. Fulvus. Biochemical characteristics of KYC 1134 broth, which showed the highest proteolytic activity, showed increased activity 8 times to seven days after culture, and protein production were increased gradually and stopped at five days. The broth had optimal temperature at $60^{\circ}C$ for proteolytic activity, and stability of pH was ranged from pH 5 to 10, at $50^{\circ}C$ and 60, respectively. To classify proteases being in the broth, ten inhibitors were determined and only bestatin showed 27% inhibition effect. The inhibition result demonstrates that the broth contains kinds of amino peptidases and other exopeptidases.
Journal of the Korea Academia-Industrial cooperation Society
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v.14
no.12
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pp.6290-6295
/
2013
This study examined the fermentative characteristics of red ginseng wine prepared with enzymatically hydrolyzed red ginseng marc (ERGM), which was produced by the polysaccharide hydrolyase and protease treatments of red ginseng marc (RGM). After adjusting the initial soluble solid contents of ERGM broth to $20^{\circ}Brix$ and $30^{\circ}Brix$ with sucrose, respectively, alcohol fermentation was carried out at $25^{\circ}C$ for 9 days. The soluble solid contents decreased from $20^{\circ}Brix$ to $7^{\circ}Brix$ and from $30^{\circ}Brix$ to $13^{\circ}Brix$, and the reducing sugar content decreased to 18 and 24 g/L at 9 days of fermentation, respectively. The ethanol contents increased to 65 g/L and 106 g/L, respectively. The crude saponin content in the ERGM broth was 2~3 times as high as that in the RGM broth after 7 days fermentation. These results show that ERGM is suitable as a raw material for the production of red ginseng wine.
Extruded defatted soy flour (DSF) with twin screw extruder was compared with steam treated DSF for soy sauce fermentation. Independent variables of response surface methodology (RSM) for extrusion were barrel temperature $(145{\sim}165^{\circ}C)$, feed moisture content $(25{\sim}35%)$, and feed rate $(20{\sim}30\;kg/hr)$. Extrusion conditions for production of the extrudates having highest water absorption capacity, lowest bulk density and highest expansion ratio were obtained by regression analysis of each dependent variable. Electron photomicrographs revealed significant increase of porosity inside the koji made from extruded DSF. Therefore, growth of fungi inside the koji made from extrudates of DSF was accelerated to achieve higher activities of protease, ${\alpha}-amylase$, and glucoamylase compared to the koji made from steamed DSF. Although amino acid composition of soy sauces prepared from two different DSF did not show any significant difference, the content of free amino acids in soluble nitrogen compounds was higher $(13%{\pm}2)$ in soy sauce made from extruded SDF. During the whole period of fermentation color of soy sauce made from extrudates of DSF was darker. Sensory evaluation did not reveal any serious off-odor and off-taste in the soy sauce fermented from extruded DSF.
Ha, Yoo Jin;Kim, Do Hyun;Lee, Byung Hee;Yoo, Sun Kyun
Journal of the Korean Applied Science and Technology
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v.35
no.2
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pp.367-377
/
2018
Swimming crab(Ovalipes punctatus) is produced in Korea and utilized as semi-processed food at streamed cooked state. Recently, protein hydrolysates have been known as having function such as antioxidant, suppression of hypertension, immunodulatory, alleviation of pain, and antimicrobial activity. This research was investigated to find the functional antioxidant from crab hydrolysates. To fine optimal protease enzyme, alcalase, bromelain, flavourzyme, neutrase, papain, and protamex were selected to evaluate the DPPH radical scavenging activity and finally bromelain to show the best activity was selected. The molecular weight of bromelain hydrolysates were distributed with range from 500 to 3,200 Da and 7 different molecules or more. The amino acids related to antioxidant capacity was about 42.54%. The processes optimization study used was the response surface methodology. The ranges of processes were the reaction temperature of 40 to $60^{\circ}C$, pH 6 to 8, and enzyme concentration 1 to 3%(w/v). As a result, the optimization of process was determined at temperature of $55^{\circ}C$, pH of 6.5, and enzyme concentration of 3%(w/v). In these conditions, degree of hydrolysates were maximum 71.60%. Therefore, we expect that those products are useful as functional food ingredients.
Wheat straw was treated on laboratory scale with 4% urea at a moisture level of 50% along with different amount of HC1 to fix various levels of ammonia (30, 40, 50 and 60%) and stored for 4 weeks. Result, revealed a significant (p<0.01) increase in CP content of the samples where HC1 was added. The CP content of the straw was only 7.8%, which increased to 14.1, 16.0, 15.0 and 15.2% with the addition of acid. Similarly the concentrations of NDF, ADF and hemicellulose was significantly different due to HC1 addition. The level of HC1 recommended was to trap 30% ammonia as there was not significant difference in CP content of straw due to addition of 4 levels of acids. Results of in vivo experiment conducted on nine buffaloes divided randomly into three groups of three animals in each revealed no significant difference in the intake of DM, OM, NDF, ADF, cellulose and hemicellulose in group I (ammoniated straw), group II (HC1 treated ammoniated straw) and group III (HC1 treated ammoniated straw + 1 kg barley grain), but the intake of CP was significantly (p<0.01) more in group III as compared to other 2 groups. The digestibility of DM, OM and CP was significantly (p<0.01) more in groups where HCI treated straw was fed as compared to only ammoniated straw fed group, whereas there was no significant difference in the digestibility of NDF, ADF and cellulose in 3 groups. Intake was significantly higher of nitrogen (p<0.05), calcium (p<0.01) and phosphorus (p<0.01) in group III as compared to other two groups. Animals in all the 3 groups showed positive nitrogen, calcium and phosphorus balance, though the balances of all the 3 nutrients were significantly higher in group III as compared to other 2 groups. Rumen fermentation study conducted in 3 rumen fistulated buffaloes in $3{\times}3$ latin square design offering the same 3 diets as in group I to III revealed that rumen pH was alike statistically in 3 groups and at various time intervals. The mean ammonia-N concentration was significantly (p<0.01) more in group II and III as compared to group I. The mean TVFA concentration (mM/100 ml SRL) were 6.46, 7.84 and 8.47 in 3 groups respectively and different statistically (p<0.01). Results revealed no significant difference in the activities of carboxy methyl cellulase, urease or protease at both the time of sampling (0 h and 4 h) in all the 3 groups of animals.
This study was conducted to investigate effects of heat-oxidized soy protein isolate (HSPI) on growth performance, serum biochemical indices, apparent nutrient digestibility and digestive function of broiler chickens. A total of 320 1-day-old Arbor Acres chicks were randomly divided into 4 groups with 8 replicates of 10 birds, receiving diets containing soy protein isolate (SPI, control group) or the same amount of SPI heated in an oven at $100^{\circ}C$ for 1, 4, or 8 h, for 21 days, respectively. The results indicated that compared with the control group, body weight gain and feed intake of birds fed diet containing SPI heated for 8 h were significantly lower (p<0.05). Serum urea nitrogen concentration was higher in the broilers fed diet containing SPI heated for 4 or 8 h at d 21 (p<0.05). In contrast, serum glucose content was decreased by HSPI substitution at d 21 (p<0.05). The relative pancreas weight in HSPI groups was higher than that in the control group at d 21 (p<0.05). Meanwhile, the opposite effect was observed for relative weight of anterior intestine and ileum in broilers fed a diet containing SPI heated for 8 h (p<0.05). Birds fed diets containing SPI heated for 4 or 8 h had a decreased lipase activity in anterior intestinal content at d 14 and 21 (p<0.05), respectively. In addition, the same effect was also noted in broilers given diets containing SPI heated for 1 h at d 21 (p<0.05). Similarly, amylase, protease and trypsin activity in anterior intestinal content were lower in broilers fed diets containing SPI heated for 8 h at d 21 (p<0.05). The apparent digestibility of dry matter (DM) from d 8 to 10 and DM, crude protein (CP), and ether extract from d 15 to 17 were lower in broilers fed diets containing SPI heated for 8 h (p<0.05). Besides, birds given diets containing SPI heated for 4 h also exhibited lower CP apparent digestibility from d 15 to 17 (p<0.05). It was concluded that HSPI inclusion can exert a negative influence on the growth performance of broilers, which was likely to result from the simultaneously compromised digestive function.
Human caspase-9, an essential apoptosis initiator protease, was excessively degraded when expressed in Escherichia coli under the conventional induction condition. To optimize the conditions for induction and develop a rapid purification method for obtaining significant amounts of wild-type procaspase-9, we expressed procaspase-9 as GST fusion in E. coli. The addition of 0.01 mM IPTG as an inducer to the bacterial culture and decreasing the culture temperature to 25oC improved the production of procasapse-9 protein by circumventing proteolytic degradation in E. coli. The wild-type procaspae-9 was purified to approximately 70% purity with relatively high yields using the method developed in this study. In addition, we found that GST-caspase-9 is autocatalytically cleaved after aspartic acid 315, which is the same site for processing in mammalian cells, during expression in E. coli.
A total of sixty-six samples of Nuruk, a fermention starter used to make the Korean traditional rice wine, Makgeolli, were collected from central and southern regions of Korea in 2013 and 2014. We classified two groups of the Nuruk samples, "commercial" and "home-made", according to the manufacturing procedure and purpose of use. Commercial Nuruks were made in a controlled environment where the temperature and humidity are fixed and the final product is supplied to Makgeolli manufacturers. Home-made Nuruks were made under uncontrolled conditions in the naturally opened environment and were intended for use in the production of small amounts of home-brewed Makgeolli. We obtained more than five hundred isolates including filamentous fungi and yeasts from the Nuruk samples followed by identification of fungal species. Also we stored glycerol stocks of each single isolate at $-70^{\circ}C$. We identified the species of each isolate based on the sequences of ITS regions amplified with two different universal primer pairs. We also performed morphological characterization of the filamentous fungi and yeast species through observations under the microscope. We investigated the major fungal species of commercial and home-made Nuruks by counting the colony forming units (CFU) and analyzing the occurrence tendency of fungal species. While commercial Nuruks contained mostly high CFU of yeasts, home-made Nuruks showed relatively high occurrence of filamentous fungi. One of the representative Nuruk manufacturers used both domestic wheat bran and imported ones, mainly from US, as raw material. Depending on the source of ingredient, the fungal diversity was somewhat different. Another commercial Nuruk sample was collected twice, once in 2013 and again in 2014, and showed different diversity of fungal species in each year. Nuruks obtained from the southern regions of Korea and Jeju island showed high frequency of yeast such as Saccharomycopsis fibuligera and Pichia species as well as unique filamentous fungus, Monascus species. S. fibuligera was easily found in many Nuruk samples with high CFU. The major filamentous fungi were Aspergillus, Lichtheimia, Mucor and Penicillium species. In order to further our understanding of the isolates and their potential industrial applications, we assayed three enzymes, alpha amylase, glucoamylase and acid protease from 140 isolates out of about five hundred isolates and selected about 10 excellent strains with high enzyme activities. With these fungal isolates, we will perform omics analyses including genomics, transcriptomics, metabolic pathway analyses, and metabolomics followed by whole genome sequencing of unique isolates associated with the basic research of Nuruk and that also has applications in the Makgeolli making process.
Yeonsan Ogae has been known as supporting health and high efficacy of treatment. In recent days, as the efficacy of functional peptides has known, the optimization of oligo peptides production and its characteristics from Ogae viscera has been performed. Response surface method was used to perform the optimizaion of enzyme hydrolysis. The range of processes was temperature (40, 50 and $60^{\circ}C$), pH(6.0, 7.0 and 8.0), and enzyme(1, 2 and 3%). The degree of hydrolysis, amono acids, molecular weight of products were analyzed. The optimum process of enzyme hydrolysis were determined as temperature $58^{\circ}C$, pH 7.5, and enzyme concetration 3%. At optimum conditions, the degree of hydrolysis after 2 h reaction was 75-80%. The total amino acids of amino acid and were 386.15 mg/100 g and 155.26 mg/100 g, respectively. The molecular weight of products by using Maldi-TOF was ranged from 300 to 1,000 Da.
The enzymatic activity of Nuruk and the quality properties of Yakju were investigated according to different fungi. The fungi that were used in this study were Aspergillus kawachii KCCM 32819, Aspergillus niger KCCM 32005, Rhizopus japonicus KCCM 11604, Rhizopus oryzae KCCM 11272, Rhizopus oryzae KCCM 11273, Rhizopus oryzae KCCM 11276, and Mucor rouxii KCCM 60148. The study results are as follows. The saccharogenic power of Rhizopus oryzae KCCM 11272 Nuruk was the highest (3,647.72 SP/g) among all the samples. The ${\alpha}$-amylase production and protease activities were highest (3.76 DU and 4.7 tyrosine mg/min, respectively) in the Rhizopus japonicus KCCM 11604 Nuruk. The pH levels of the Yakju made with commercial Nuruk and Rhizopus japonicus KCCM 11604 Nuruk were 4.14 and 4.07, respectively. The total titratable acid content of the Yakju made with Rhizopus oryzae KCCM 11273 Nuruk was the highest (0.56%) among all the samples. Rhizopus japonicus KCCM 11604 and Rhizopus oryzae KCCM 11272 had the highest ethanol yields (15.18% and 15.10%, respectively). In the sensory evaluation carried out in this study, the panel preferred the Yakju made with Rhizopus japonicus KCCM 11604 Nuruk. Overall, however, the panel did not like the Yakju made with Aspergillus niger KCCM 32005 Nuruk.
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