• Journal of Microbiology and Biotechnology
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• v.1 no.2
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• pp.136-141
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• 1991

Optimum culture conditions for the formation of nitrile hydratase by Brevibacterium sp. CH2 were investigated. Addition of ferric and ferrous ions greatly increased the nitrile hydratase formation. The effects of nitriles, amides, and acids as an inducer on the formation of nitrile hydratase were investigated. Isobutyramide was the best inducer among the tested compounds. When Brevibacterium sp. CH2 was cultivated for 23 h at $30^{\circ}C$ in a optimized medium containing 15 g of glucose, 5 g of bacto peptone, 3 g of yeast extract, 3 g of malt extract, 1 g of $KH_2$$PO_4$, 1 g of $K_2$$HPO_4$, 1 g of NaCl, 0.5 g of isobutyramide, 0.2 g of MgSO$_4$ㆍ7$H_2O$, and 0.02g of $FeSO_4$ㆍ$7H_2$O per liter of distilled water with pH controlled at 7.1, the maximum total activity was 665 units/ml of the culture broth and the specific activity was 70 units/mg of the dry cells. The medium optimization increased the specific activity of Brevibacterium sp. CH2 2.2 times.

• Journal of Microbiology
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• v.44 no.4
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• pp.439-446
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• 2006

In this study, the growth kinetics of Lactobacillus rhamnosus and lactic acid production in continuous culture were assessed at a range of dilution rates $(0.05 h^{-1}\;to\;0.40h^{-1})$ using a 2L stirred tank fermenter with a working volume of 600ml. Unstructured models, predicated on the Monod and Luedeking-Piret equations, were employed to simulate the growth of the bacterium, glucose consumption, and lactic acid production at different dilution rates in continuous cultures. The maximum specific growth rate of L. rhamnosus, ${\mu}_{max}$, was estimated at $0.40h^{-1}$I, and the Monod cell growth saturation constant, Ks, at approximately 0.25g/L. Maximum cell viability $(1.3{\times}10^{10}CFU/ml)$ was achieved in the dilution rate range of $D=0.28h^{-1}\;to\;0.35h^{-1}$. Both maximum viable cell yield and productivity were achieved at $D=0.35h^{-1}$. The continuous cultivation of L. rhamnosus at $D=0.35h^{-1}$ resulted in substantial improvements in cell productivity, of 267% (viable cell count) that achieved via batch cultivation.

• KSBB Journal
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• v.17 no.4
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• pp.326-332
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• 2002

The quality of biopharmaceutical proteins is strongly affected by a manufacturing process employed to produce Et, and thus validation of the manufacturing bioprocess is a very important issue. Chromatography is probably the most widely used bioprocess unit operation for protein purification. In this study, a miniaturized automatic chromatography system was designed and constructed for scale-down studies for process chromatography validation. This system, named MiniValChrom, has the following features: automatic and repeated operation, flexible sequences and intervals among the steps, on-line and real-time monitoring and control, method files savings, etc. Using the MiniValChrom, we peformed a case study of an abbreviated experiment to estimate chromatographic resin lifetime. BSA (bovine serum albumin) and Cibacron Blue 3G-A were used as the model protein and the resin, respectively. The resin deterioration was evaluated by determining and monitoring the HETP and NTP values from the chromatograms every 5 cycles. It was observed that the HETP and the NTP values were changed by 9% after 15 cycles. The resin lifetime validation could be completed by repeating this experiment until the HETP value reached a predetermined value. The MiniValChrom's concept and the protocol suggested in this study can serve as a rapid and economical tool for the validation studies of bioprocess chromatography system.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.1
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• pp.72-79
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• 2006

Kojic acid production by Aspergillus flavus strain S44-1 using sucrose as a carbon source was carried out in a 250-mL shake flask and a 2-L stirred tank fermenter. For comparison, production of kojic acid using glucose, fructose and its mixture was also carried out. Kojic acid production in shake flask fermentation was 25.8 g/L using glucose as the sole carbon source, 23.6 g/L with sucrose, and 6.4 g/L from fructose. Reduced kojic acid production (13.5 g/L) was observed when a combination of glucose and fructose was used as a carbon source. The highest production of kojic acid (40.2 g/L) was obtained from 150 g/L sucrose in a 2 L fermenter, while the lowest kojic acid production (10.3 g/L) was seen in fermentation using fructose as the sole carbon source. The experimental data from batch fermentation and resuspended cell system was analysed in order to form the basis for a kinetic model of the process. An unstructured model based on logistic and Luedeking-Piret equations was found suitable to describe the growth, substrate consumption, and efficiency of kojic acid production by A. flavus in batch fermentation using sucrose. From this model, it was found that kojic acid production by A. flavus was not a growth-associated process. Fermentation without pH control (from an initial culture pH of 3.0) showed higher kojic acid production than single-phase pH-controlled fermentation (pH 2.5, 2.75, and 3.0).

• Microbiology and Biotechnology Letters
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• v.38 no.1
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• pp.7-12
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• 2010

The biochemical study of sugar uptake in yeasts started five decades ago and led to the early production of abundant kinetic and mechanistic data. However, the first accurate overview of the underlying sugar transporter genes was obtained relatively late, due mainly to the genetic complexity of hexose uptake in the model yeast, Saccharomyces cerevisiae. The genomic era generated in turn a massive amount of information, allowing the identification of a multitude of putative sugar transporter and sensor-encoding genes in yeast genomes, many of which are phylogenetically related. This review aims to briefly summarize our current knowledges on the biochemical and molecular features of the transporters of pentoses in yeasts, when possible establishing links between previous kinetic studies and genomic data currently available. Emphasis is given to recent developments concerning the identification of D-xylose transporter genes, which are thought to be key players in the optimization of S. cerevisiae for bioethanol production from lignocellulose hydrolysates.

• Korean Journal of Microbiology
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• v.52 no.1
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• pp.18-24
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• 2016

We investigated 23 lactic acid bacteria isolated from Korean breast milk-fed infant in order to select strains which show superior anti-allergic effect. The candidates were cultivated and then we obtained dried powders of tyndallized cells and supernatant concentrate separately. Screening was carried out with down-regulation of interleukin (IL) 4 and up-regulation of IFN-${\gamma}$ in mouse splenocytes. As a result of the screening, we selected Lactobacillus rhamnosus IDCC 3201 (RH3201) for oral feeding to ovalbumin-sensitized BALB/c mice. Oral administration of RH3201 as dead cell bodies and supernatant concentrate suppressed hyper-production of serum immunoglobulin (Ig) E levels compared to vehicle group. Such anti-allergic effects were achieved by improvement of the balance between cytokines produced from type-1 helper T (Th1) and type-2 helper T (Th2) lymphocytes. Therefore, RH3201 has potential to improve atopic symptoms by immunomodulatory effect.

• Journal of Microbiology and Biotechnology
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• v.20 no.5
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• pp.946-949
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• 2010

The effects of two different sugars (glucose and xylose) on the expression levels and patterns of the xylose reductase (xyl1), xylitol dehydrogenase (xyl2), and xylulokinase (xyl3) genes were analyzed using Pichia stipitis. A significant increase in mRNA levels of xyl1 was observed after 6 h growth in culture conditions using xylose as a sole carbon source, but expressions of the three genes were not influenced by normal culture media with glucose. In addition, expressions of xyl2 and xyl3 were not observed during the entire culture period during which xylose was added. It also was found that the expression level of xyl1 increased as a function of the xylose concentration (40, 60, and 80 g/l) used in this study, indicating that xyl1 expression sensitively responded to xylose in the culture media. Although the induced level of xyl2 increased slightly after 48 h in the xylose-supplemented culture conditions, the expression of xyl2 was not observed in the xylitol-supplemented culture conditions. Finally, considering the expression of each gene in response to glucose or xylose, the absolute expression levels of the three genes indicate that xyl1 is induced primarily by exposure to xylose.

• Microbiology and Biotechnology Letters
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• v.49 no.2
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• pp.181-191
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• 2021

The present study addresses isolation, optimization, partial purification, and characterization of a haloalkaline serine protease from a newly isolated haloarchaeal strain isolated from Wadi El Natrun in Egypt. We expected that a two-step sequential statistical approach (one variable at a time, followed by response surface methodology) might maximize the production of the haloalkaline serine protease. The enzyme was partially purified using Hiprep 16/60 sephacryl S-100 HR gel filtration column. Molecular identification revealed the newly isolated haloarchaeon to be Natrialba hulunbeirensis strain WNHS14. Among several tested physicochemical determinants, casamino acids, KCl, and NaCl showed the most significant effects on enzyme production as determined from results of the One-Variable-At-A-time (OVAT) study. The BoxBehnken design localized the optimal levels of the three key determinants; casamino acids, KCl, and NaCl to be 0.5% (w/v), 0.02% (w/v), and 15% (w/v), respectively, obtaining 62.9 U/ml as the maximal amount of protease produced after treatment at 40℃, and pH 9 for 9 days with 6-fold enhancement in yield. The enzyme was partially purified after size exclusion chromatography with specific activity, purification fold, and yield of 1282.63 U/mg, 8.9, and 23%, respectively. The enzyme showed its maximal activity at pH, temperature, and NaCl concentration optima of 10, 75℃, and 2 M, respectively. Phenylmethylsulfonyl fluoride (PMSF, 5 mM) completely inhibited enzyme activity.

• Proceedings of the Korean Aquaculture Society Conference
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• 2006.05a
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• pp.83-84
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• 2006

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.59-60
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• 2003