• Food Industry And Nutrition
• /
• v.18 no.1
• /
• pp.8-12
• /
• 2013

• Bulletin of Food Technology
• /
• v.24 no.4
• /
• pp.525-543
• /
• 2011

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

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.5 no.4
• /
• pp.275-287
• /
• 2000

Fermentation strategies for recombinant protein production in Pichia pastoris have been investigated and are reviewed here. Characteristics of the expression system, such as phenotypes and carbon utilization, are summarized. Recently reported results such as growth model establishment, app58lication of a methanol sensor, optimization of substrate feeding strategy, DOstat controller design, mixed feed technology, and perfusion and continuous culture are discussed in detail.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.4 no.1
• /
• pp.36-40
• /
• 1999

Optimal lyophilization process was developed for manufacturing the dried product of Lactobacillus acidophilus with high cell viability. Three major factors, freezing rate, specific surface area of samples, and stabilizer type and their synergy were shown to play a crucial role in the development of an effective lyophilization process. Finally we found an optimal combination among three process parameters mentioned above; an exceptionally high cell survival percentage of 90% was achieved using the 8.28 cm-1 specific surface area of samples, slow freezing rate, and a stabilizer composition of 4% skim milk +1% glycerol +0.1% calcium chloride.

• Journal of Microbiology and Biotechnology
• /
• v.1 no.3
• /
• pp.182-187
• /
• 1991

The effects of acrylonitrile and acrylamide on the enzyme action of nitrile hydratase of Brevibacterium sp. CH1 and CH2 strains used for the biotransformations of nitriles were studied. The excessive substrate (acrylonitrile) and product (acrylamide) inhibited the enzyme activity competitively. In comparison with 0.2 mol/l of CH1 strain, the substrate inhibition of CH2 strain began to appear only at a high acrylonitrile concentration of 0.91 mol/l. In a packed bed reactor, dispersed plug flow model was proposed and this model was proved to be valid by the experiment. Also acrylamide productivity decreased sharply when acrylamide concentration in the substrate solution exceeded 20％ (wt/v).

• KSBB Journal
• /
• v.15 no.5
• /
• pp.423-427
• /
• 2000

Intense research on biosensors has been performed in a number of different institution over the past 15 years, but relatively few commercial products have resultingly, the blood glucose sensor is a good example of a product which penetrated the market. However recently, the development of electrochemical and optical technologies has accelerated the turnover of the research as is illustrated by a rapid increase in the number of point-of-care diagnostic systems and analytical devices. Examples of such biosensors used in the fields of medical diagnostics, bioprocess control, and environmental monitoring are described, and summarized in an introduction to their characteristics, structures, and functions, given.

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

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

• 한국생물공학회:학술대회논문집
• /
• 2001.11a
• /
• pp.274-277
• /
• 2001

This research was conducted as a part of continuous development of a bioprocess to treat thiocyanate($SCN^-$) in the leachate from an industrial landfill site. Detailed characteristics of the leachate, physical and chemical, was analyzed in this study. Time to reach equilibrium in $SCN^-$ degradation at different temperature, similar to the annual variation. was investigated for posssible adaptation of the bioprocess in the treatment site. Complete mineralization of $SCN^-$ to ammonia. bicarbonate and sulfate at all temperature range with except $5^{\circ}C$ trial was observed with different time elapsed to reach equilibrium. which shortened as the process of Fill-Draw proceeded. The final metabolites were mainly nitrate and sulfate. No fouling odor was detected and the settling characteristic of the biomass after the treatment was good(i.e., sludge volume index(SVI) ${\le}$80). Using MWT sludge as an inoculum showed better performance with respect to the elapsed time for treatment and sedimentation characteristics than those of using 2BET sludge.