• KSBB Journal
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• v.30 no.6
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• pp.307-312
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• 2015

Transgenic rice cells using RAmy3D promoter can provide high productivity, and the production of recombinant protein is induced by sugar starvation. In this system, productivity was reduced during the scale-up processes. To ensure the influences of shear stress and oxygen transfer rate, working volume and mixing performances were investigated under various agitation speeds and working volumes. In addition, inoculation methods including suspended cells and filtered cells were compared. Working volumes and shaking speeds were 300, 450 mL and 80, 120 rpm, respectively. Hydrodynamic environment of each condition was measured numerically like mixing time and $k_La$. Good mixing performance and high shear stress were measured at high agitation speed and low volume. The highest level of hCTLA4Ig was 30.7 mg/L at 120 rpm, 300 mL. When conditioned medium was used for inoculation, increased cell growth was noticed during the day 0~4 and decreased slower than filtered cells. Compared with filtered cells, the maximum hCTLA4Ig level reached 37.8 mg/L at 120 rpm, 300 mL and lower protease activity level was observed. In conclusion mixing performance is critical factor for productivity and conditioned medium can have a positive effect on damaged cells caused by hydrodynamic shear stress.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.532.2-532
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• 1986

Alkaline protease which is an important enzyme used in detergents, leather tanning and food industry was produced by alkalophilic bacterium, Bacillus sp. KCTC 1723 isolated from soil. The maximum productivity of the enzyme in alkaline medium containing 1% sodium bicarbonate was obtained by incubating for 3 days at 37$^{\circ}C$. The optimum pH of the enzyme was 11.5 and calcium ion was effective on stabilization of the enzyme at high temperature. The enzyme was not inhibited by metal chelating agent such as El)TA but inhibited by diisopropyl fluorophosphate. Purification of the enzyme was carried out DEAE- and CM-cellulose column chromatographies and molecular weight of the purified enzyme was determined

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

• The Korean Journal of Food And Nutrition
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• v.7 no.1
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• pp.8-15
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• 1994

To utilize hemicellulosic biomass efficiently, the microorganism producing xylanase was isolated from fermented sawdust. It was a Gram positive, aerobic and rod shape bacterium. It had endospore and secreted strong hydrolases, such as amylase and protease. Through morphological, cultural and physiological tests, it was identified as Bacillus sp. GS. To increase the productivity of xylanase from Bacillus sp. GS, the enzyme production medium was optimized. The composition of the medium and incubation conditions were like follows xylan 1.25%, yeast extract 0.1%, NaN030.2%, K2HP04 0.1%, MgSO4 0.02%, mineral salt 0.005%, pH 6.5, incubation temperature 37$^{\circ}C$.

• Korean Journal of Food Science and Technology
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• v.42 no.1
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• pp.39-44
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• 2010

The process of the preparation of branched-chain amino acid (BCAA)-enriched hydrolysates from corn gluten was optimized through the parameters of pre-treatment (heating and cellulosic hydrolysis), hydrolysis method (acid, protease, and microbe plus protease), concentration, and spray drying condition. The protein yield of corn gluten was increased by heating and cellulase treatments. Among three different hydrolysis methods, the combined use of microbes and protease was the most effective in terms of free amino acid (FAA) and BCAA content of the corn gluten hydrolysates. In addition, the FAA and BCAA content in the hydrolysates prepared by microbial and enzymatic combined treatment were improved by a concentration process. Spray drying conditions for the preparation of the powder from the hydrolyzed reactant were an inlet temperature of $185^{\circ}C$, outlet temperature of $80^{\circ}C$, and the use of maltodextrin as an anticaking agent. Thus, this study established an economical process for preparation of value-added hydrolysates of excellent productivity and quality, in terms of high BCAA content and product stability.

• Journal of Microbiology and Biotechnology
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• v.9 no.3
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• pp.282-291
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• 1999

Lactic acid bacteria were isolated from Kimchi and screened for bacteriocin. A total of 99 strains showed antimicrobial activity when grown on solid media, yet only 10 showed antimicrobial activity in liquid media. Strain H-559, identified as Lactococcus lactis subsp. lactis, exhibited the strongest inhibitory activity and was active against pathogenic bacteria including Listeria monocytogenes, Staphylococcus aureus, and Bacillus cereus as well as other lactic acid bacteria. The antimicrobial substance produced by L. lactis subsp. lactis H-559 was confirmed to be a bacteriocin by the treatment of $\alpha$-chymotrypsin, and protease type Ⅸ and ⅩIV. The bacteriocin activity remained stable between pH 2.0 and pH 11.0 and during heating for 10 min at $100^{\circ}C$. The bacteriocin production started in the exponential phase and stopped in the stationary phase. L. lactis subsp. lactis H-559 showed the highest bacteriocin activity at a culture temperature of $25^{\circ}C$, and an inverse relationship between the bacteriocin productivity and mean growth rate at different culture temperatures was observed. The mean growth rate and bacteriocin productivity of L. lactis subsp. lactis H-559 increased as the initial pH of the media increased.

• Research in Plant Disease
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• v.22 no.2
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• pp.81-93
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• 2016

Out of plant-associated bacteria, certain plant growth-promoting bacteria (PGPB) have been reported to increase plant growth and productivity and to elicit induced resistance against plant pathogens. In this study, our objective was to broaden the range of applications of leaf-colonizing PGPB for foliar parts of road tress and pepper. Total 1,056 isolates of endospore-forming bacteria from tree phylloplanes were collected and evaluated for the enzymatic activities including protease, lipase, and chitinase and antifungal capacities against two fungal pathogens, Colletotrichum graminicola and Botrytis cinerea. Fourteen isolates classified as members of the bacilli group displayed the capacity to colonize pepper leaves after spraying inoculation. Three strains, 5B6, 8D4, and 8G12, and the mixtures were employed to evaluate growth promotion, yield increase and defence responses under field condition. Additionally, foliar application of bacterial preparation was applied to the road tress in Yuseong, Daejeon, South Korea, resulted in increase of chlorophyll contents and leaf thickness, compared with non-treated control. The foliar application of microbial preparation reduced brown shot-hole disease of Prunus serrulata L. and advanced leaf abscission in Ginkgo biloba L. Collectively, our results suggest that leaf-colonizing bacteria provide potential microbial agents to increase the performance of woody plants such as tree and pepper through spray application.

• Korean Journal of Food Science and Technology
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• v.22 no.6
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• pp.611-617
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• 1990

This study was conducted to breed a high vitamin $B_{12}$ producer by the fusion of protoplasts between Bacillus natto and Bacillus megaterium. Auxotrophic mutants of Bacillus natto SH-34 ($thr^-try^-rif^r$) and Bacillus megaterium BK-13 ($arg^-ade^-lys^-str^r$) which showed high protease activity and production of vitamin $B_{12}$, respectively, were isolated for the fusion experiment. Protoplasts were induced by incubating the cells with lysis solution containing $500{\mu}/ml$ lysozyme, and the ratio of protoplast and regeneration formation were ranged from 99% and 67%, respectively. Fusion frequencies of fusants between Bacillus natto SH-34 and Bacillus megaterium BK-13 were appeared in the ranges of $1.0{\times}10^{-5}$ under the treatment of 30% PEG 6000 containing 3% PVP. The fusant, MNF-72 showed the highest product yield of $7.85{\mu}g/g-cell\;vitamin\;B_{12}$ in production medium. For the improvement of productivity, the immobilization of fusants with sodium alginate was carried out. In batch and continuous fermentation systems, the productivity were determined to be $0.58{\mu}g/ml.hr\;and\;0.80{\mu}g/ml.hr\;vitamin\;B_{12}$ under optimum condition, respectivity.

• The Plant Pathology Journal
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• v.33 no.5
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• pp.458-466
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• 2017

Xanthomonas oryzae pv. oryzae (Xoo), the causative agent of bacterial blight, is a major threat to rice productivity. Here, we performed RNA-Seq based transcriptomic analysis of Xoo transcripts isolated under in planta growth (on both susceptible and resistant hosts) and in vitro culture conditions. Our in planta extraction method resulted in successful enrichment of Xoo cells and provided RNA samples of high quality. A total of 4,619 differentially expressed genes were identified between in planta and in vitro growth conditions. The majority of the differentially expressed genes identified under in planta growth conditions were related to the nutrient transport, protease activity, stress tolerance, and pathogenicity. Among them, over 1,300 differentially expressed genes were determined to be secretory, including 184 putative type III effectors that may be involved in Xoo pathogenicity. Expression pattern of some of these identified genes were further validated by semi-quantitative RT-PCR. Taken together, these results provide a transcriptome overview of Xoo under in planta and in vitro growth conditions with a focus on its pathogenic processes, deepening our understanding of the behavior and pathogenicity of Xoo.

• The Plant Pathology Journal
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• v.37 no.6
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• pp.632-640
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• 2021

Cucumber mosaic virus (CMV) and Pepper mild mottle virus (PMMoV) causes severe economic loss in crop productivity of both agriculture and horticulture crops in Korea. The previous surveys showed that naturally available biopolymer material - chitosan (CS), which is from shrimp cells, reduced CMV accumulation on pepper. To improve the antiviral activity of CS, it was synthesized to form phosphate cross-linked chitosan (PCS) and compared with the original CS. Initially, the activity of CS and PCS (0.01%, 0.05%, and 0.1% concentration) compound against PMMoV infection and replication was tested using a half-leaf assay on Nicotiana glutinosa leaves. The total number of local lesions represented on a leaf of N. glutinosa were counted and analyzed with phosphate buffer treated leaves as a negative control. The leaves treated with a 0.1% concentration of CS or PCS compounds exhibited an inhibition effect by 40-75% compared with the control leaves. The same treatment significantly reduced about 40% CMV accumulation measured by double antibody sandwich enzyme-linked immunosorbent assay and increased the relative expression levels of the NPR1, PR-1, cysteine protease inhibitor gene, LOX, PAL, SRC2, CRF3 and ERF4 genes analyzed by quantitative reverse transcriptase-polymerase chain reaction, in chili pepper plants.