It is necessry to develop and strengthen the activity of enzymic source which in low applied for maggerley brewing as an amylolytic and proteolytic starter, recently in this country the active and strong enzymic starter is required for the better brewing and to substitute another starch material for the present wheat flour. In this study, manufacturing method the strong enzymic source have been developed and established with use of raw wheat bran plus fungal strains of Rhizopus sp. and Aspergillus usamii the culture of starter. The results on experimental the activities of enzymic sources (stater) are as following ; 1. Method of making the enzymic source (starter) is to cultivate the strains of Asperguillus orzyae, Asp. kawachii, Asp usamii and Rhizopus sp. in the acid treated raw or heatboiled wheat bran. 2. The saccharogenic pwoer (S.P.) of enzymic source which consisted of raw bran plus fungi and cultured in it is generally stronger than those of heat-boiled bran plus fungi, the strongest power was shown in the culture of Rhizopus plus raw bran, and the next other is in mixture of Asp.usamii and Rhizopus on raw wheat bran. 3. The most strong alpha amylase activity was expressed in the plot of Asp.oryzae on heat-boiled wheat bran, the next was in the culture of Rhizopus nad Aspergillus usamii on raw wheatbran. 4. The most vigourous acidic proteinase activity was expressed in the micture of raw bran plus Asp. usamii and Rhizopus those were independentlu cu;tured before mixing for neutral proteinase activity, it was shown in the mixed culture of Asp. usamii and Rhizopus on raw wheat bran, the msot active alkaline proteinase activity of enzymic source was found in the plot of raw bran material. 5. For poly-preptidase activity in pH 6.5 it is found that the culture of Rhizopus and Asp.usamii on raw bran was most active among them of enzymic sources. 6. Generally, it is concluded that culture of fungi on acid treated raw wheat bran is stronger in its activity than those of heat boiled wheat bran, especially the culture of Rhizopus nad Asp.usamii on raw bran exhibited the most vigorous and non-polarized activity for all aspects, so it is considered to be most desirable enzymic stater in Korean Maggerley brewing and this would be able to substitute brewing material for the present wheat flour because of its strong and wide hand activity of amylolytic and proteolytic action.
An expression vector system was developed for the secretory production of recombinant Bacillus stearothermophilus L1 lipase in Saccharomyces cerevisiae. The mature L1 lipase gene was fused to ${\alpha}-amylase$ signal sequence from Aspergillus oryzae for the effective secretion into the culture broth and the expression was controlled under GAL10 (the gene coding UDP-galactose epimerase of S. cerevisiae) promoter. S. cerevisiae harboring the resulting plasmid successfully secreted L1 lipase into the culture broth. To examine an optimum condition for L1 lipase expression in the fed-batch culture, L1 lipase expression was induced at three different growth phases (early, mid, and late-exponential growth phases). Maximum product on of L1 lipase (1,254,000 U/l, corresponding to 0.65/1) was found when the culture was induced at an early growth phase. Secreted recombinant L1 lipase was purified only through CM-Sepharose chromatography, and the purified enzyme showed 1,963 U/mg of specific activity and thermoalkalophilic properties similar to those reported for the enzyme expressed in Escherichia coli.
This study was conducted to restore our traditional Cheongtaejeon tea and to develop the special products. We inoculated Cheongtaejeon tea with lactobacilli (Lactobacillus plantarum CHO 25) and the mixed microbial strains (L. plantarum CHO 25 + Saccharomyces cerevisiae + Bacillus amyloliquefaciens CHO 104). We also examined the sensuous characteristics and physiological activity of Cheongtaejeon tea which was produced by the inoculation of microbial strains. The external appearance of Cheongtaejeon teas were not significant among the teas which were produced with or without the inoculation of L. plantarum CHO 25 and the mixed microbial strains. The taste of the tea increased most in Cheongtaejeon tea which was produced without the inoculation of microbial strains. The taste and liking of Cheongtaejeon tea which was inoculated with Aspergillus oryzae and Aspergillus niger decreased significantly, and it was not suitable to drink. Total phenolics compound contents, total flavonoid contents and DPPH ($\alpha,\alpha$-diphenyl-$\beta$-picryl-hydrazyl) radical scavenging activity of Cheongtaejeon tea extracts increased much more in the order of that produced with the inoculation of L. plantarum CHO 25, control and that with the mixed microbial strains. However, nitrite radical scavenging activity in 1,000 mg/L Cheongtaejeon tea hot water extracts were in the order of the control (94.4%), the inoculation of L. plantarum CHO 25 (93.6%) and the mixed microbial strains (91.1%). Overall results indicated that the sensuous characteristics increased most in Cheongtaejeon tea which was produced without the inoculation of microbial strains, and those physiological activities in tea with the inoculation of L. plantarum CHO 25.
The in vivo effect of tyrosinase inhibitors in the melanogenesis of gold fish (jet black color) was evaluated by measuring surface color and observing melanin pigment. The fish was firstly cultivated in 0.9% NaCl solution for 1 week to induce melanogenesis, and then, it was transferred to each treatment group containing tyrosinase inhibitor. The fish was grouped into control. food additive group (addition of 5 mM glutathione, 5 mM cysteine, and 1 mM benzoic acid), microbial inhibitor group (addition of culture broth of Aspergillus oryzae in shiitake and glucose medium), and plant extract group (addition of the mixed extracts of green tea, beet, red chicory, and nameko). After 6 days, the fish was anesthetized by electric shock, and color of pectoral region, lateral region, and dorsal fin was measured. Hunter's L and b values of treated group were generally higher than those of control group, indicating that the tyrosinase inhibitors could inhibit the melanogenesis of the fish. Effect of plant extract was apparent, though relatively weak, not because it did not work in vivo, but because a sufficient amount of extract could not be added to fish globes. If a large amount of extract was added, fish gradually died due to a microbial contamination. Microscopic observation of melanin in lateral scale and dorsal fin showed that in the treated groups with tyrosinase inhibitors, the number of melanophore per unit area and the size of one melanophore decreased.
Zinc is an essential trace element in Human Being. Highly zinc containing yeast strain isolated from the tropical fruit, rambutan and the zinc concentration in this yeast strain was 306 ppm (30.6 mg%)per dry matter basis. This strain was found to be a rounded type, normal size, and multi-polar budding. Phylogenetic analysis using the ITS1-5.85 rDNA sequences from isolated strain is most similar to yeast Saccharomyces cerevisiae at the level of nucleotide sequence identity at 99%. This strain was produced alcohol by about 12% using fully colonized koji-rice with Aspergillus oryzae. In conclusion, the isolated strain was found to be closely related to the S. cerevisiae based on its morphological and physiological properties, and alcohol fermentation. The phylogenetic analysis of strain FF-10 using ITS 5.8S rDNA sequence data also supported the closely related to the S. cerevisiae. Accordingly, the isolated yeast was named as S. cerevisiae FF-10. Further studies on the best culture conditions for zinc production from zinc-enriched S. cerevisiae FF-10 are under investigation.
In the search for novel potent fungi-derived bioactive compounds for bioinsecticide applications, crude ethyl acetate culture filtrate extracts from 110 mangrove fungal endophytes were screened for their toxicity. Toxicity tests of all extracts against brine shrimp (Artemia salina) larvae were performed. The extracts with the highest toxicity were further examined for insecticidal activity against Spodoptera litura larvae and acetylcholinesterase (AChE) inhibition activity. The results showed that the extracts of five isolates exhibited the highest toxicity to brine shrimp at 50% lethal concentration ($LC_{50}$) values of 7.45 to 10.24 ppm. These five fungal isolates that obtained from Rhizophora mucronata were identified based on sequence data analysis of the internal transcribed spacer region of rDNA as Aspergillus oryzae (strain BPPTCC 6036), Emericella nidulans (strains BPPTCC 6035 and BPPTCC 6038), A. tamarii (strain BPPTCC 6037), and A. versicolor (strain BPPTCC 6039). The mean percentage of S. litura larval mortality following topical application of the five extracts ranged from 16.7% to 43.3%. In the AChE inhibition assay, the inhibition rates of the five extracts ranged from 40.7% to 48.9%, while eserine (positive control) had an inhibition rate of 96.8%, at a concentration of 100 ppm. The extracts used were crude extracts, so their potential as sources of AChE inhibition compounds makes them likely candidates as neurotoxins. The high-performance liquid chromatography profiles of the five extracts differed, indicating variations in their chemical constituents. This study highlights the potential of culture filtrate ethyl acetate extracts of mangrove fungal endophytes as a source of new potential bioactive compounds for bioinsecticide applications.
Song, Sang Hoon;Lee, Chunghee;Lee, Sulhee;Park, Jung Min;Lee, Hyong-Joo;Bai, Dong-Hoon;Yoon, Sung-Sik;Choi, Jun Bong;Park, Young-Seo
Journal of Microbiology and Biotechnology
/
v.23
no.1
/
pp.40-46
/
2013
A variety of nuruk were collected from various provinces in Korea, and their microflora profiles were analyzed at the species level. A total of 42 nuruk samples were collected and when the viable cell numbers in these nuruk were enumerated, the average cell numbers of bacteria, fungi, yeast, and lactic acid bacteria from all nuruk were 7.21, 7.91, 3.49, and 4.88 log CFU/10 g, respectively. There were no significant differences in viable cell numbers of bacteria or fungi according to regions collected. Bacillus amyloliquefaciens and B. subtilis were the predominant bacterial strains in most samples. A significant portion, 13 out of 42 nuruk, contained foodborne pathogens such as B. cereus or Cronobacter sakazakii. There were various species of lactic acid bacteria such as Enterococcus faecium and Pediococcus pentosaceus in nuruk. It was unexpectedly found that only 13 among the 42 nuruk samples contained Aspergillus oryzae, the representative saccharifying fungi in makgeolli, whereas a fungi Lichtheimia corymbifera was widely distributed in nuruk. It was also found that Pichia jadinii was the predominant yeast strain in most nuruk, but the representative alcohol fermentation strain, Saccharomyces cerevisiae, was isolated from only 18 out of the 42 nuruk. These results suggested that a variety of species of fungi and yeast were distributed in nuruk and involved in the fermentation of makgeolli. In this study, a total of 64 bacterial species, 39 fugal species, and 15 yeast species were identified from nuruk. Among these strains, 37 bacterial species, 20 fungal species, and 8 yeast species were distributed less than 0.1%.
Bamboo leaf powder was added to Chiffon cake to increase its neutraceutical effects and storage periods. Specifically, 0, 7, 10 and 13% leaf powder was added to the cakes. The content of total dietary fiber in the bamboo leaf powder was 65.57%. The moisture content did not differ significantly among groups. The cake containing 10% bamboo leaf powder had the greatest volume, whereas the control group (0% bamboo leaf powder) had the lowest volume (p<0.05). The Hunter's L and a values decreased significantly as the amount of bamboo leaf powder increased. The b value of the control was lowest among the groups (p<0.05). Evaluation of the consumer acceptance of flavor revealed that the cakes containing the added bamboo leaf had greater consumer acceptance than the control. However, when the color was evaluated, the 13% group showed the lowest acceptance (p<0.05). Other factors such as texture, taste and overall acceptance did not differ significantly among groups. Additionally, the elasticity did not differ among groups, while the air cells were most uniform in the control. The strength of bamboo leaf aroma, bitter taste and aftertaste increased as the amount of bamboo leaf powder added increased. Cakes containing 10% and 13% added powder had the greatest moisture content, while the control had the lowest content (p<0.05). As in previous studies, the results of this study indicated that 10% bamboo leaf powder was the optimal level for the preparation of Chiffon cake. To evaluate the storage of cakes, the 10% group and the control were inoculated with Aspergillus oryzae and then incubated at $30^{\circ}C$ for 6 days. The microbial colony counts in the control group were dramatically increased after 48 hrs; however, the fungal concentration of the 10% group did not increase for 4 days. In conclusion, the addition of 10% bamboo leaf powder to Chiffon cake increased the storage time while maintaining adequate consumer acceptance.
The quality characteristics of soy yogurt prepared with different proteolytic enzymes and starter culture were evaluated. In order to facilitate the growth of lactic acid bacteria and subsequent production of lactic acid, soy protein isolate(SPI) was hydrolyzed using three kinds of proteases; one extracted from Aspergillus oryzae, bromelain and ${\alpha}-chymotrypsin$. The cultural systems employed thereafter for lactic fermentations were: 1) Bifidobacterium bifidum, 2) B. bifidum and Lactobacillus acidophilus, 3) B. bifidum and Lactobacillus bulgaricus. In soy yogurt, pH was more decreased by mixed culture method than single culture method with the accumulation of lactic acid. Viable cells of lactic acid bacteria in soy yogurts were measured $10^8$ CFU/g by the single culture method while $10^9$ CFU/g by the mixed culture method except ${\alpha}-chymotrypsin$ treatment. The amount of free amino acids in soy yogurts were substaintially increased by enzyme treatment. Viscosity was decreased by enzyme treatment, resulting in higher viscosity by ${\alpha}-chymotrypsin$ treatment. Water holding capacity was found to be higher in the single culture method in case of enzyme treatment. Among the various volatile flavor components isolated and identified after enzyme hydrolysis, acetaldehyde, ethanol, diacetyl, butyl alcohol contents tended to increase by lactic fermentation.
A strain of Streptomyces sp producing chitinase was isolated from soil and its cultural condition and some properties of this enzyme were investigated. When 0.375 per cent of glucose was added to basal medium, this organism produced the most quantities of this enzyme after shaking culture at 3$0^{\circ}C$ for 48 hrs., while the production of the enzyme was repressed at the more concentration of glucose than that. The enzyme had a optimal pH of 7.0, optimal temperature of 5$0^{\circ}C$ and the activity of that was not decreased by heat treatment for 20 minute at 7$0^{\circ}C$. And then the activity was increased by Co$^{2+}$ but was slightly inhibited by Hg$^{2+}$, Ni$^{2+}$, Pb$^{2+}$.EX> 2+/.
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