• Journal of Microbiology and Biotechnology
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• v.24 no.1
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• pp.52-58
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• 2014

Structurally, herboxidiene contains the tetrahydropyran acetic acid moiety and a side chain including a conjugated diene, and has been isolated from Streptomyces chromofuscus ATCC 49982. Its production was significantly elevated nearly 13.5-fold (0.74 g/l) in a medium supplemented with glycerol (medium No. 6A6), and was more efficacious (1.08 g/l; 19.8-fold) in fed-batch fermentation at 36 h in medium No. 6A6, from Streptomyces chromofuscus. For further enhancement, regulatory genes metK1-sp and afsR-sp from Streptomyces peucetius were overexpressed using an expression vector, pIBR25, and similarly ACCase from Streptomyces coelicolor and two genes, metK1-sp and afsR-sp, were also overexpressed using an integration vector, pSET152, under the control of the strong $ermE^*$ promoter in Streptomyces chromofuscus. Only the recombinant strains S. chromofuscus SIBR, S. chromofuscus GIBR, and S. chromofuscus AFS produced more herboxidiene than the parental strain in optimized medium No. 6A6 with an increment of 1.32-fold (0.976 g/l), 3.85-fold (2.849 g/l), and 1.7-fold(1.258 g/l) respectively.

• KSBB Journal
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• v.10 no.4
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• pp.455-460
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• 1995

Specific growth rate was controlled for the repression of acetic acid formation in the fed-batch fermentation of recombinant Escherichia coli. With controlled specific growth rate, we studied the effect of the specific growth rate on cell growth, glucose consumption, acetic acid formation, and the expression of recombinant protein (${\beta}$-lactamase). High specific growth rate caused the accumulation of glucose and acetic acid, and lowered the production of recombinant protein. However, the addition of methionine recovered the gene expression by alleviating the negative effect of acetic acid at high specific growth rate.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.3
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• pp.196-200
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• 2004

The supra molecular weight poly(〔R〕-3-hydroxybutyrate) (PH B), having a molecular weight greater than 2 million Da, has recently been found to possess improved mechanical properties compared with the normal molecular weight PHB, which has a molecular weight of less than 1 million Da. However, applications for this PHB have been hampered due to the difficulty of its production. Reported here, is the development of a new metabolically engineered Escherichia coli strain and its fermentation for high level production of supra molecular weight PHB. Recombinant E. coli strains, harboring plasm ids of different copy numbers containing the Alcaligenes latus PHB biosynthesis genes, were cultured and the molecular weights of the accumulated PHB were compared. When the recombinant E. coli XL1-Blue, harboring a medium-copy-number pJC2 containing the A. latus PHB biosynthesis genes, was cultivated by fed-batch culture at pH 6.0, supra molecular weight PHB could be produced at up to 89.8 g/L with a productivity of 2.07 g PHB/L-h. The molecular weight of PHB obtained under these conditions was as high as 22 MDa, exceeding by an order of magnitude the molecular weight of PHB typically produced in Ralstonia eutropha or recombinant E. coli.

• BMB Reports
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• v.38 no.3
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• pp.294-299
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• 2005

In this report, we describe the high-yield secretory expression of the recombinant human anti-HBsAg Fab fragment from Pichia pastoris that was achieved by co-integration of the genes encoding the heavy and light chains (both under the control of alcohol oxidase promoter) into the genome of the yeast cells. The fed-batch fermentations were carried out in a 5 L scale. Both chains of the Fab were successfully expressed upon methanol induction. The absorbance ($OD_{600}$) of the broth can reach 350~500 at the end of fed-batch phase. After the induction, the expression level of the recombinant Fab (soluble) reached 420~458 mg/L. The recombinant Fab fragment was purified from the crude culture supernatant by ion exchange chromatography and the purity of the recombinant Fab fragment was over 95%. The affinity activities of the crude fermentation supernatant and the purified Fab were analyzed by indirect ELISA, which showed that the purified recombinant Fab fragment had high affinity activity with hepatitis B surface antigen.

• Korean Journal of Agricultural Science
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• v.41 no.3
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• pp.237-243
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• 2014

Human Glucagon like peptide-1 (GLP-1) is an incretin hormone that promotes secretion of insulin. In order to eliminate the formation of the soluble aggregate, Ala19 in GLP-1 was substituted with Thr, resulting in a GLP-1 mutant GLP-1A19T. The gene synthesis of GLP-1A19T and the fusion of 6-lysine tagged ubiquitin gene were accomplished by using the overlap extension polymerase chain reaction. The ubiquitin fused GLP-1A19T (K6UbGLP-1A19T) is expressed as form of inclusion body with little formation of the soluble aggregation in recombinant E. coli. In order to produce K6UbGLP-1A19T in large amounts, fed-batch fermentation was carried out in a pH-stat feeding strategy. Maximum dry cell weight of 87.7 g/L and 20.4% of specific K6UbGLP-1A19T content were obtained. Solid-phase refolding using a cation exchanger was carried out to renature K6UbGLP-1A19T. The refolded K6UbGLP-1A19T aggregated little and was released GLP-1A19T by on-column cleavage with ubiquitin-specific protease-1. The molecular mass of GLP-1A19T showed an accurate agreement with its theoretical molecular mass.

• KSBB Journal
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• v.18 no.1
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• pp.14-18
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• 2003

In this study of the simultaneous saccharification and fermentation (SSF) of paper sludge, fed-batch cultivation of Lactobacillus paracasei subsp. paracasei KLB58 was attempted to produce viable KLB58 cells and lactic acid. Optimal culture conditions, including the temperature and concentration of the supplemented enzyme, were examined in terms of lactic acid production and viable cell count. When the effects of culture temperature and $\beta$-glucosidase concentration were examined in fed-batch SSF, the highest viable cell counts and lactic acid production (i.e. 5$\times$$10^9$ CFU/ml and 45 g/L, respectively) were obtained at 37$^{\circ}C$ and 2 unit/ml of $\beta$-glucosidase.

• Journal of Microbiology and Biotechnology
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• v.22 no.7
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• pp.967-974
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• 2012

Cultivation of Spirulina platensis using ammonium salts or wastewater containing ammonium as alternative nitrogen sources is considered as a commercial way to reduce the production cost. In this research, by analyzing the relationship between biomass production and ammonium-N consumption in the fed-batch culture of Spirulina platensis using ammonium bicarbonate as a nitrogen nutrient source, an online adaptive control strategy based on optical density (OD) measurements for controlling ammonium feeding was presented. The ammonium concentration was successfully controlled between the cell growth inhibitory and limiting concentrations using this OD-based feedback feeding method. As a result, the maximum biomass concentration (2.98 g/l), productivity (0.237 g/l d), nitrogen-to-cell conversion factor (7.32 gX/gN), and contents of protein (64.1%) and chlorophyll (13.4mg/g) obtained by using the OD-based feedback feeding method were higher than those using the constant and variable feeding methods. The OD-based feedback feeding method could be recognized as an applicable way to control ammonium feeding and a benefit for Spirulina platensis cultivations.

• KSBB Journal
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• v.5 no.1
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• pp.25-35
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• 1990

In the Xanthan gum fermentation by Xanthomonas campestris there are problems of the large energy consumption by long fermentation time, the mass transfer of oxygen and nutrients by high viscous fermentation broth. In this study, the media optimization and the fed batch fermentation were carried out to decrease fermentation time and increase Xanthan gum yield. The $O_2$ uptake rate (OUR) and $CO_2$ evolution rate(CER) which were obtained from the analysis of fermentation exit gas using a gas chromatograph were investigated. As a result, the fermentation time decreased at optimal assimilable nitrogen concentration but increased at poor or rich assimilable nitrogen concentration, the Xanthan gum biosynthesis was stimulated under the limited condition of assimilable nitrogen source and the optimum fermentation medium was obtained as follow; Glucose=30g / l, Peptone=8.0g / l, $K_2HPO_4=2.0g/l$, $MgS0_47H_2O=10g/l$, Sodium acetate=20g/l, Sodium pyruvate=0.5g/1. As the agitation speed and nitrogen concentration increased, the $O_2$ uptake rate and $CO_2$ evolution rate increased. The OUR and CER were 37.3mmol $O_2/\;l$ hr and 20.2 mmol $CO_2/\;L$ hr at peptone 11g / l and agitation speed 990RPM, respectively. In fed batch fermentation, the final concentration of Xanthan gum was enhanced up to 29g / l.

• Journal of Microbiology and Biotechnology
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• v.34 no.9
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• pp.1912-1918
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• 2024

Single-cell protein (SCP) derived from microorganisms is widely recognized as a viable alternative protein source for the future. Nevertheless, the commercialization of yeast-based SCP is hampered by its relatively low protein content. Therefore, this study aimed to enhance the protein content of Saccharomyces cerevisiae via random mutagenesis. To achieve this, S. cerevisiae KCCM 51811, which exhibited the highest protein concentration among 20 edible S. cerevisiae strains, was selected as a chassis strain. Subsequently, a KCCM 51811 mutant library was constructed (through UV irradiation) and screened to isolate mutants exhibiting high protein content and/or concentration. Among the 174 mutant strains studied, the #126 mutant exhibited a remarkable 43% and 36% higher protein content and concentration, respectively, compared to the parental strain. Finally, the #126 mutant was cultured in a fed-batch system using molasses and corn-steep liquor, resulting in a protein concentration of 21.6 g/l in 100 h, which was 18% higher than that produced by the parental strain. These findings underscore the potential of our approach for the cost-effective production of food-grade SCP.

• International Journal of Industrial Entomology and Biomaterials
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• v.6 no.1
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• pp.15-19
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• 2003

The study evaluated the influence of feeding different maturity leaves i.e., tender, medium, coarse and mixed leaves of three different maturity during late larval stages of new bivoltine race $(CSR_{3} {\times} CSR_{6})$. The data were compared with shoot feeding and control batches in which conventional feeding method was followed. The most of the larval and cocoon characters were recorded significantly higher in tender leaves fed batches followed by T4 batch (2 times tender and 1 time coarse leaves). Lowest melting percentage (1.494%) was recorded in T4 and highest (4.69%) was recorded in coarse leaf (T3) fed batches. Significantly higher post cocoon parameters viz., average filament length, non-breakable filament length, renditta and raw silk percentage were recorded in tender loaves fed batches.