• Journal of Microbiology and Biotechnology
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• v.7 no.4
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• pp.282-286
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• 1997

The floc strength of Escherichia coli was enhanced by adding water soluble polymer flocculants (BPA-5020 and BPA-5000) to the particulate flocculant (BPA-1000) as indicated by the increase in the shear index. The shear index of the E. coli flocs increased from 0.39 with the particulate flocculant alone to 0.94 with the particulate flocculant in conjunction with the water soluble polymer flocculant. In addition, the sedimentation rate of flocs was higher and the sedimented volume of flocs was smaller when the particulate flocculant was used with the water soluble polymer flocculant. When E. coli was flocculated first with the water soluble flocculant and the particulate flocculant was added later into the E. coli flocs formed, the sedimentation rate of the flocs was greater than that of any other combination. The shear index of the flocs was, however, independent of the sequence of the flocculant addition.

• Journal of Applied Biological Chemistry
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• v.48 no.4
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• pp.167-169
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• 2005

Crude extract of bioconverted linoleic acid using Pseudomonas aeruginosa PR3 was evaluated for its antibacterial activity against food-borne pathogenic bacteria. Crude extract showed antibacterial activity against four Gram-positive bacteria, Staphylococcus aureus (ATCC 6538), S. aureus (KCTC 1916), Listeria monocytogenes (ATCC 19166), and Bacillus subtilis (ATCC 6633), and one Gramnegative bacterium, Pseudomonas aeruginosa (KCTC 2004), with minimum inhibitory concentrations ranging from 750 to $1,500\;{\mu}g{\cdot}ml^{-1}$. S. aureus and B. subtilis were selected for growth inhibition assays with bioconverted linoleic acid. Major antibacterial effects occurred at lag phase.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.1
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• pp.84-87
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• 2006

Batch binding experiments were performed to assess the recovery performance of glyceraldehyde 3-phosphate dehydrogenase (G3PDH) bound to the unshielded and polymer (polyvinyl pyrrolidone. PVP)-shielded dye-ligand (Cibacron Blue 3GA) adsorbent. The adoption of a polymer-shielded, dye-ligand technique facilitated the elution efficiency of bound G3PDH. It was demonstrated that the recovery of G3PDH using polymer-shielded dye-ligand adsorption yielded higher elution efficiency, at 60.5% and a specific activity of 42.3 IU/mg, after a low ionic strength elution (0.15 M NaCl). The unshielded dye-ligand yielded lower elution efficiency. at 6.5% and a specific activity of 10.2 IU/mg.

• Journal of Microbiology and Biotechnology
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• v.18 no.5
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• pp.889-896
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• 2008

A suitable simple model tested by experiments is required to address complex biological reactions like esterase synthesis by Saccharomyces cerevisiae. Such an approach might be the answer to a proper bioprocessing strategy. In this regard, a logistic model for esterase production from Saccharomyces cerevisiae has been developed, which predicts well the cell mass, the carbon source (glucose) consumption, and the esterase activity. The accuracy of the model has been statistically examined by using the Student's t-test. The parameter sensitivity analysis showed that all five parameters (${\mu}_m$, $K_a$, $X_m$, $Y_{x/s}$, and $Y_{p/x}$) have significant influence on the predicted values of esterase activity.

• Food Science and Industry
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• v.52 no.2
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• pp.171-187
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• 2019

Novel food materials can be produced based on biotechnology such as genetic recombination, microbial fermentation, and enzymatic engineering by utilizing living organisms such as animal, plant, and microorganism or by applying the enzymes isolated from them. Especially, exploration and development of novel prebiotics and probiotics attracted great attention worldwide in the food industry, of which the research and industrial trends in food biotechnology field are promoting the production of next generation sweeteners and proliferation of beneficial bacteria in gastrointestinal tract. Development and commercialization of novel food materials by domestic bioprocessing technology have been sluggish due to the GMO/LMO food safety issues. Meanwhile, the US and EU do not perceive badly about gene manipulation technology, and the research is most active in the fields of crops and GMMs, respectively. Genetic scissors, which are considered as next generation technology, are notable since foreign genes do not remain in final products.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.2
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• pp.208-213
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• 2007

The objective of the experiment was to determine the optimum cultural [moisture levels (55, 60 and 70%), days of fermentation (7, 14 and 21), temperature (25 and $35^{\circ}C$) of incubation)] and nutritional parameters (urea addition (0 and 2%) and variable levels of single super phosphate (0.25 and 0.50% SSP)) for bio-processing of the mustard (Brassica campestris) straw (MS) under solid-state fermentation (SSF) system. The performance of SSF was assessed in terms of favorable changes in cell wall constituents, protein content and in vitro DM digestibility of the MS. Sorghum based inoculum (seed culture) of Ganoderma lucidum to treat the MS was prepared. The 50 g DM of MS taken in autoclavable polypropylene bags was mixed with a pre-calculated amount of water and the particular nutrient in the straw to attained the desired levels of water and nutrient concentration in the substrate. A significant progressive increase in biodegradation of DM (p<0.001), NDF (p<0.01) and ADF (p<0.05) was observed with increasing levels of moisture. Among the cell wall constituents the loss of ADF fraction was greatest compared to that of NDF. The loss of DM increased progressively as the fermentation proceeded and maximum DM losses occurred at 28 days after incubation. The protein content of the treated MS samples increased linearly up to the day $21^{th}$ of the incubation and thereafter declined at day $28^{th}$, whereas the improvement in in vitro DM digestibility were apparent only up to the day $14^{th}$ of the incubation under SSF and there after it declined. The acid detergent lignin (ADL) degradation was slower during the first 7 days of SSF and thereafter increased progressively and maximum ADL losses were observed at the day $28^{th}$ of the SSF. The biodegradation of DM and ADL was not affected by the variation in incubation temperature. Addition of urea was found to have inhibitory effect on fungal growth. The effect of both the levels (0.25 and 0.50) of SSP addition in the substrate, on DM, NDF, ADF, cellulose and ADL biodegradation was similar. Similarly, the protein content and the in vitro DM digestibility remain unaffected affected due to variable levels of the SSP inclusion in the substrate. From the results it may be concluded that the incubation of MS with 60 percent moisture for 21 days at $35^{\circ}C$ with 0.25 percent SSP was most suitable for MS treatment with Ganoderma lucidum. Maximum delignification, enrichment in the protein content and improvement in in vitro DM digestibility were achieved by adopting this protocol of bioprocessing of MS.

• KSBB Journal
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• v.21 no.5
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• pp.360-365
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• 2006

The objectives of this study were to develop nano-pore silica particles and to modify its surface for use as an enzyme immobilization matrix. Sol-gel reaction was used to produce silica particles of various nano pore sizes with hydroxyl groups on their surfaces. The surface was modified with aldehyde that was confirmed by fluorescence imaging. Trypsin was covalently immobilized by reductive amination. Surface density of the immobilized trypsin was ca. $350{\mu}g/m^2$, which was approximately 17- and 35-fold higher than those from the surfaces with hydroxyl and amine group, respectively. About 90% of the initial enzyme activity was maintained after the 12th use of repeated use. When compared with the commercial matrices, the nano-pore silica particle was superior in terms of immobilization yield and specific activity. This study suggests the nano porous silica particles can be used as enzyme immobilization matrix for industrial applications.

• KSBB Journal
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• v.14 no.6
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• pp.651-654
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• 1999

Using recombinant human growth hormone as a model protein, we carried out unfolding by adding a denaturant such as urea, guanidine HCl, or SDS followed by refolding by dilution and dialysis. The objectives were to monitor the structural changes during in vitro refolding process and, based on the results, to develop a quantitative method of refolding progress assessment. The changes in surface hydrophobicity were measured by fluorescence tagging of 1-anilinonaphthalene-8-sulfonate(1,8-ANS) to the hydrophobic portions, and those in the secondary structure were monitored by using far UV-CD(circular dichroism) spectroscopy. Also, we used RP-HPLC to separate and quantify the folded and unfolded proteins to correlate the result with the structure analysis. Our results indicate the surface hydrophobicity are well correlated with the formations of the secondary structure, primarily ${\alpha}$-helices, as well as the disulfide bridges. We expect this monitoring technique can be applied in industrial fields as a means to quantitatively assess the progress of in-vitro refolding of recombinant proteins.

• KSBB Journal
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• v.21 no.2
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• pp.133-139
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• 2006

In 'solid-phase' PEGylation, the conjugation reaction occurs as the proteins are attached to a solid matrix, and thus it can have distinct advantages over the conventional, solution-phase process. We report a case study: rhIFN-${\alpha}$-2a was first adsorbed to cation exchange resin and then N-terminally PEGylated by aldehyde mPEG of 5, 10, and 20 kD through reductive alkylation. After the PEGylation, salt gradient elution efficiently recovered the mono-PEGylate in a purified form from the unwanted species such as unmodified IFN, unreacted PEG, and others. The mono-PEGylation and its purification were integrated in a single chromatographic step. Depending on the molecular weight of the mPEG aldehyde used, the mono-PEGylation yield ranged 50-64%. We could overcome the major problems of random, or uncontrollable, multi-PEGylation and the post-PEGylation purification difficulties associated with the solution-phase process. N-terminal sequencing and MALDI-TOF MS confirmed that a PEG molecule was conjugated only to the N-terminus. Compared with the unmodified IFN, the mono-PEGylate showed the reduced anti-viral activity as measured by the cell proliferation assay. The bioactivity was reduced more as the higher molecular weight PEG was conjugated. Immunoreactivity, evaluated indirectly by antibody binding activity using a surface plasmon resonance biosensor, also decreased. Nevertheless, trypsin resistance as well as thermal stability was considerably improved.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.375-382
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• 2010

The available techniques for heterologous protein secretion in Lactobacillus strains are limited. The aim of the present study was to develop an efficient protein-secretion system using recombinant lactobacilli for various applications such as live delivery of biotherapeutics. For the construction of expression vectors, the Lactobacillus brevis slpA promoter, Lactobacillus casei prtP signal sequence, and mouse IL-10 sequences were used as a model system. Interestingly, the slpA promoter exhibited strong activity in L. casei, contrary to previous observations. In order to stabilize replication of the plasmid in E. coli, a removable terminator sequence was built into the promoter region. For the improvement of secretion efficiency, a DTNSD oligopeptide was added to the cleavage site of signal peptidase. The resulting plasmids provided remarkably efficient IL-10 secretion. Accumulation of the protein in the culture supernatant varied widely according to the pH conditions. By analysis of the secreted protein, formation of homodimers, and biological activity, IL-10 was confirmed to be functional. The presently constructed plasmids could be useful tools for heterologous protein secretion in L. casei.