• Microbiology and Biotechnology Letters
• /
• v.38 no.4
• /
• pp.379-384
• /
• 2010

We isolated microorganisms presenting high enzymatic activities for amylase, cellulase, protease, lipase or fibrinolysis from Korean traditional soybean sauce and paste. Then, the physiological properties and 16S rRNA sequences of isolated microorganisms were analyzed. All of the isolated 13 strains possessing high extra cellular enzyme activities have higher amylase and cellulase activities than Bacillus subtilis KACC 10114. All the selected strains have protease activities except for D2-14. Except D8-8 and K4-1, other strains have lipase activity. D2-7, D8-8 and K4-1 strains have higher fibrinolytic activities than others, while D8-2 strain has no activity. Most of the selected strains showed antibacterial activity even in gram positive and gram negative bacteria and yeast. Gene sequence analysis of 16S rRNA from isolated strains revealed that all the selected strains were member of Bacillus species.

• Applied Biological Chemistry
• /
• v.14 no.3
• /
• pp.191-200
• /
• 1971

As a series on the soy-bean protein and their related substances 9 samples were collected from 9 places such as straws (Rice) to obtain bacterial strains which produce protease. From these samples total of 23 strains were isolated by the use of dilution pour plate method. For all isolated strains primary screening of productivity of protease was performed and useful straines with regard to protease productivities were identified. Optimum conditions for enzyme action of protease from isolates $D_9$, $F_{20}$ strains were pH 7.5 and $40^{\circ}C$. Chung-Kook-Jang is one of the characteristic foods in Korea made from soy-bean by fermentation. The chief bacterium is Bacillus subtilis and the chief change which takes place in soy-bean during fermentation is degradation of protein. Three kinds of Chung-Kook-Jang were prepared using three different strains of Bacillus natto, $D_9\;and\;F_{20}$ from isolated. Water soluble-N, TCA soluble-N, amino-N and peptide-N were measured about the steamed soybean, Chung-Kook-Jang prepared with three strains of bacteria. Water soluble-N decreased very largely in steamed soybean, but in Chung-Kook-Jang it increased to 85% of raw soy-bean.

• Korean Journal of Microbiology
• /
• v.49 no.1
• /
• pp.38-45
• /
• 2013

In this study, euryhaline marine microorganism, Bacillus sp. strain EBW4 isolated from polychaete (Perinereis aibuhitensis) of Suncheon Bay was physiologically, biochemically and genetically characterized. Based on 16S rRNA sequence, EBW14 was found to share 98.25% similarity with Bacillus hemicentroti $JSM076093^T$, 97.96% similarity with Bacillus hwajinponensis SW-$72^T$ and 96.28% similarity with B. algicoa $KMM3737^T$, respectively. The temperature range for the growth of strain EBW4 was $4-40^{\circ}C$, NaCl concentration range 0-17% and pH range pH 5-9, revealing that EBW4 was euryhaline bacterium. Major fatty acids in strain EBW4 were composed of anteiso $C_{15:0}$ (48.2%), iso $C_{16:0}$ (12.1%), anteiso $C_{17:0}$ (11.6%) and iso $C_{14:0}$ (9.4%). EBW4 was found to have DNase, amylase, protease and lipase for the degradation of macromolecules such as DNA, carbohydrates, proteins, lipids, etc. The enzyme activities of alkaline phosphatase, esterase (C4), leucine arylamidase and ${\alpha}$-chymotrypsin were also found in strain EBW4. Analysis of the biodegradation ability of EBW4 for organic hydrocarbons under different salinity conditions using synthetic water waste revealed that EBW4 exhibited the ability to degrade organic hydrocarbons very quickly, suggesting strain EBW4 may be a good candidate for the application to various industries.

• Journal of Chitin and Chitosan
• /
• v.23 no.4
• /
• pp.285-292
• /
• 2018

In this study, for the purpose of decomposing food waste, the strain was screened from traditional fermented food and soils. The enzyme activity (protease, amylase, cellulase, lipase) experiment was carried out using the paper disc method in 212 strains isolated from 5% NaCl media. Among them, only the strains having enzyme activity of more than 2 (soil) or more than 4 (traditional fermented food) with the halozone of enzyme activity of 15 mm or more were selected first, and microorganism identification through 16S rRNA sequencing was performed. Finally, were identified such as Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus siamensis, Bacillus licheniformis, Bacillus aquimaris, Bacillus megaterium, Bacillus koreensis, Bacillus stratoshericus, Bacillus aryabhattai, Bacillus safensis, Marinobacter hydrocarbonoclasticus. 11 species of mixed strains were confirmed that the culture time was 24 hours, the incubation temperature was $30^{\circ}C$ and the optimum pH was 7.0. In order to confirm the degree of decomposition of standard food wastes (100 g) by treating 11 kinds of mixed strains (25%), solid content of more than $2000{\mu}m$ was determined to be 103 g for the sterilized water group and 18 g for the mixed strains group. And the rest was decomposed to a size of less than $2000{\mu}m$.

• Korean Journal of Microbiology
• /
• v.37 no.3
• /
• pp.214-220
• /
• 2001

To develop the effective composting system, we isolated bacteria that have the abilities to degrade organic matters such as cellulose, carbohydrate, protein and lipid during the compositing of livestock manureAmong 24 strains, 6 bacteria have all the enzymatic activities of protease, amylase, cellulase and lipase.These microorganisms were identified as Corynebacterium varibilis, Bacillus spp., Pseudomonas spinosa,Acetobacter calcoaceticus and Athrobacter cumminsii All the enzymes produced by the bacteria showedactivities at the broad pH range and the maximal activities were obtained at $60^{\circ}C$. It seemed that after theincrease of temperature caused by fermentation of livestock manure, the enzymes started to degrade the rawmaterials, which are added for the control of humidity. However cellulase activity was maximum at $37^{\circ}C$,suggesting that the cellulase-producing bacteria work at an early stage of livestock manure fermentation toprovide the organic material for the growth of other bacteria. The production of the enzymes were growth-associated and maximal activities appeared at the early stationary phase of growth.

• Microbiology and Biotechnology Letters
• /
• v.10 no.4
• /
• pp.275-281
• /
• 1982

One strain of Streptomyces sp. (AS-707) isolated from soil was found to produce a biologically active substance that showed a strong inhibitory activity against proteolytic enzymes viz. trypsin, papain, $\alpha$-chymotrypsin, Azotobacter protease, and Bacillus pretense. The substance was separated from culture filtrate by ion exchange column chromatography using Amberlite IRC-50 and CM-cellulose column chromatography. It was found that the recovery yield was 26% as activity basis. The substance was stable in wide pH range from 2.0 to 12.0 at 37$^{\circ}C$, but it was unstable in alkaline pH values at 6$0^{\circ}C$. The activity was thermostable to give 90% activity compared to the intact sample when it was treated at pH5.6 at 10$0^{\circ}C$ for 2 hours.

• Animal Bioscience
• /
• v.35 no.10
• /
• pp.1628-1634
• /
• 2022

Objective: This study focused on the effect of dietary metallo-protease and Bacillus velezensis CE 100 on growth performance, carcass parameters, intestinal microflora, footpad dermatitis (FPD), and manure odor in broiler chickens. Methods: One hundred-ten (two-day-old Ross 308) broiler chicks were randomly assigned to five groups with two replicate pens. The dietary treatments were divided to control, metallo-protease groups (A1, added with 0.1%; A2, added with 0.2%) and B. velezensis CE 100 groups (B1, added with 0.5%; B2, added with 1.0%). Results: The feed intake was decreased in A1 and B2 compared to the other group (p<0.05). The liver weight was lower in B2 than in A2 (p<0.05). The Salmonella in the cecum was decreased in A2 compared to control and A1 (p<0.05). However, the lactic acid bacteria were increased in all treatments (p<0.05). The litter moisture content was decreased in A2, B1, and B2 (p<0.05). The litter quality visual score was increased in all treatments (p<0.05). The FPD score and prevalence were reduced in all treatments (p<0.05). The (CH₃)₂S emission was decreased in all treatments (p<0.05). Conclusion: The present study indicated that both additives improve litter quality and reduce the incidence of FPD. These findings suggest that dietary metallo-protease and B. velezensis CE 100 have the potential to improve the broiler chickens' welfare.

• Journal of Life Science
• /
• v.11 no.4
• /
• pp.304-310
• /
• 2001

In order to produce a high quality and functional black bean Chungkugjang, a bacterium which has potent enzyme activities(protease:124.8 U/$m\ell$, $\alpha$-amylase: 78.2U/$m\ell$, glucoamylase; 13.9U/$m\ell$, ), was isolated by using the halo zone method and identified by morphological, cultural and biochemical properties, and then finally confirmed by GP-microplate identification system as Bacillus megatrium SMY-212. The isolated SMY-212 system was also high in the formation of mucoid material and fibrinolytic activity.

• Microbiology and Biotechnology Letters
• /
• v.23 no.5
• /
• pp.544-549
• /
• 1995

The alkaline elastase is an extracellular serine protease of the alkalophilic Bacillus strain Ya-B. To increase the gene copy number and the production level of the alkaline elastase Ya-B, we designed, on the B. subtilis chromosome, a gene amplification of the 10.6 kb repeating unit containing amyE, aleE (alkaline elastase Ya-B gene) and tmrB. The aleE was inserted between amyE and tmrB, and B. subtilis APT119 strain was transformed with this amyE-aleE-tmrB-junction region fragment. As a result, we succeeded in obtaining tunicamycin-resistant (Tm$^{r}$) transformants (Tf-1, Tf-2) in which the designed gene amplification of 10.6 kb occurred in chromosome. The transformants showed high productivity of $\alpha $-amylase and alkaline elastase Ya-B. The copy number of the repeating unit (amyE-aleE-tmrB) was estimated to be 25, but plasmid vector (pUC19) was not integrated. The amplified aleE of chromosome was more stable than that of plasmid in absence of antibiotics.

• Korean Journal of Microbiology
• /
• v.30 no.1
• /
• pp.60-64
• /
• 1992

The secretion vector with promoter and signal sequence region of neutral protease gene (npr) from Bacillus amyloliquefaciens was constructed by the technique of polymerase chain reaction (PCR). A unique restriction iste was introduced into the 3' of the signal coding region by the synthesis of PCR primer. To demonstrate the function of cloned promoter and signal sequence, we used the E. coli .betha.-lactamase structural gene as a foreign gene. The signal sequence of .betha.-lactamase gene was deleted by Bal31 exonuclease and only mature region was introduced into the secretion vector. Bacillus subtilis cells transformed by the recombinant vector synthesized the fusion protein and were also capable of removing the signal peptide from the original fusion protein, as judged by the assay of .betha.-lactamase activity and secretion into the growth medium by western blotting.