• Journal of Life Science
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• v.10 no.4
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• pp.397-402
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• 2000

Polyphosphate kinase catalyzes the formation of polyphosphate from ATP. To understand the mechanism of phosphate accumulation, the Serratia marcescens gene encoding ppk was cloned from the genomic library by the method of Southern hybridization. The hybridization positive DNA fragment region from pDH3 was subcloned into the expression vector. The ppk gene product, a polypeptide of 75 kDa, was confirmed by SDS-PAGE. Expression of the Serratia marcescens ppk is regulated by the catabolite repression system. The enzyme activity polyphosphate kinase was increased in the E. coli strain harboring plasmid pMH4 with ppk gene.

• Journal of Plant Biotechnology
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• v.31 no.2
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• pp.175-178
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• 2004

Transgenic tobacco (Nicotiana tabacum L. cv Bright Yellow-2) cell lines expressing a human histone H1.5 (referred to as hH1.5), which suppress collagen-induced rheumatoid arthritis, were developed under the oxidative stress-inducible peroxidase (SWPA2) promoter. Tobacco BY-2 cells were transformed by Agrobacterium-mediated method. The kanamycin-resistant calli were selected on the modified MS medium containing 150mg/L kanamycin and 300mg/L claforan. Transgenic cell lines were confirmed by PCR and northern blot analysis. Recombinant hH1.5 (rhH1.5) protein (42 kDa) was also detected by Western blot analysis, showing a different molecular weight of human hH1.5 (32 kDa). These results suggested that a hH1.5 gene was properly introduced in tobacco cultured cells under the control of SWPA2 promoter. The further characterization of rhH1.5 protein remains to be studied.

• Journal of Applied Biological Chemistry
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• v.62 no.4
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• pp.361-366
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• 2019

Isorhamnetin 3-O-glucoside, a member of the flavonol group, has been reported to be effective for inflammatory and ulcer, as well as to alleviate diabetic complications such as neuropathy, nephropathy and retinopathy. Isorhamnetin 3-O-glucoside has been extracted from several plants. Biotransformation is a valuable tool, which is used to produce value-added chemicals with inexpensive compounds. To synthesis isorhamnetin 3-O-glucoside from quercetin, two genes (PGT E82L and ROMT-9) were introduced into Escherichia coli, respectively. In order to synthesis isorhamnetin 3-O-glucoside from quercetin, a co-culture fermentation system was developed by optimizing the medium and temperature for biotransformation, the cell mix ratio, Isopropyl-β-ᴅ-thiogalactoside induction time, and quercetin feed concentration. Finally, isorhamnetin 3-O-glucoside was biosynthesized up to 181.2 mg/L under the optimized biotransformation condition, which was higher 4.7 times than previously reported (39.6 mg/L).

• Journal of Food Hygiene and Safety
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• v.35 no.4
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• pp.304-311
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• 2020

Galacto-oligosaccharides (GOS) are prebiotics that have a beneficial effect on human health by promoting the growth of probiotic bacteria in the gut, in addition to having various applications in the food industry. GOS are generally produced from lactose in a reaction catalyzed by β-galactosidase. Synthesis of GOS from whey permeate (WP) (ultrafiltration of whey, concentrated then spray dried) using surface engineered β-galactosidase in Pichia pastoris (P. pastoris) is a novel method to convert waste into a valuable product. Cell-surface display is the expression of peptides and proteins on the surface of living cells by fusing them to functional components of cells. Surface engineered cells have many potential uses. The Flo1p flocculation functional domain, thought to be located near the N terminus, recognizes and adheres non-covalently to cell-wall components such as α-mannan carbohydrates, causing reversible aggregation of cells into flocs.

• Journal of Life Science
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• v.8 no.2
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• pp.181-188
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• 1998

Two distinct ADP-ribosyltransferases, termed Yac-1 and Yac-2 from mouse lymphoma cells were recently cloned and characterized. Yac-1 enzyme possesses ADP-ribosyltransferases activity. In contrast, Yac-2 has significant NAD glycohydrolase activity and may preferentially hydrolyze NAD. Yac-2 possesses a glutamate at position 219 adjacent to the two consdrved glutamic acid residues. To study the effect of Glu-219 on enzyme activities, Glu-219 was mutagenized to Glutamine (E219Q) or alanine (E219A) using a two-step recombinant polymerase chain reaction procedure. Replacing Glu at position 219 with Gln or Ala resulted in 56 (E219Q) or 66% (E219A) reduction in ADP-ribosyltranferase activity. The NAD glycohydrolase activity of Yac-2 protein were not altered by the mutations. These results indicate that Glu-219 in Yac-2 enzyme plays an important role in ADP-ribosyltransferase, but not NAD glycohydrolase activity.

• KSBB Journal
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• v.18 no.6
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• pp.500-505
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• 2003

‘LK (lipoprotein kringle) 68’is a polypeptide of a modified ansiostatin consisting of three kringle structures that might be clinically useful as a potential cancer therapeutics. It can be produced by overexpressing it as inclusion body in recombinant E. coli. In this study, solid-phase refolding processes using packed bed adsorption (PBA) and expanded bed adsorption (EBA) column were carried out to compare their refolding yields with that of the conventional, solution-phase refolding process, For the solution-phase and the PBA-mediated processes employing Q-Sepharose, washed inclusion body was used as the starting material, whereas both washed inclusion body and E. coli homogenate were used for the EBA-mediated process employing streamline DEAE. On the final recovery LK68 per unit mass of wet cell basis, the EBA- and PBA-mediated processes showed about 2.7- and 1.5-fold higher yields, respectively, than the solution-phase refolding method. The solid-phase refolded LK68 demonstrated the same Iysine binding bioactivity and the retention time in the RP-and SEC-HPLC as those of the native protein.

• KSBB Journal
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• v.9 no.3
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• pp.339-345
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• 1994

Deletions between UASG and the GALI TATA box reduced glucose repression and allowed constitutive expression of the gene product in the absence of galactose. The relative inducer level (ratio of galactose/glucose concentrations) affected the extent of gene expression and glucose repression. Glucose repression was reduced by a factor of 2 to 5 as the relative inducer level increased. In the medium containing galactose only, induction of ${\beta}$-galactosidase synthesis by galactose increased with plasmid copy number. On the contrary, plasmid copy number did not affect significantly ${\beta}$-galactosidase synthesis in the medium containing both glucose and galactose (2% glucose＋2% galactose), which might be due to glucose repression caused by high glucose concentration. However, when the medium contained the relatively high inducer level (0.4% glucose＋0.8% galactose), ${\beta}$-galactosidase synthesis increased with plasmid copy number, indicating that the beneficial effect of higher galactose concentration was weaker than the repressive effect of higher glucose concentration.

• Journal of Korean Society of Environmental Engineers
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• v.32 no.8
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• pp.795-801
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• 2010

The acute toxicity of arsenic compounds was assessed and compared using following four bioassays; bioluminescence activity of the recombinant strain RB1436, germination of four different seeds, ${\alpha}$-glucosidase activity produced by Bacillus lichemiformis, acute genetic revertant mutation using mutant strain Salmonella typhimurium. Different sensitivities were observed among tested bioassays, but generally the toxicity by arsenite was greater than that of arsenate. Among tested four seeds, sensitivities of Lactucus and Raphanus were greater than others, and these two seed types were appeared as proper type for bioassay. High revertant mutation ratio (5.1) was observed with 1 mg/L arsenite, indicating high mutagenicity. The sensitivity of ${\alpha}$-glucosidase activity on arsenic compounds was much lower than other methods. The evaluation of interactive toxic effects using various bioassays may comprise a useful tool for the bioassessment of environmental pollutants.

• Microbiology and Biotechnology Letters
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• v.13 no.4
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• pp.349-354
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• 1985

A 4.7 kb Hind III fragment containing $\alpha$-amylase gene of Bacillus stearothermophilus IAM 11062 was cloned in Escherichia coil HB101, using plasmid pBR322 and runaway plasmid pSY343 as a vector. The cloned gene was stably maintained and expressed In E.coli. The constructed strain of E. coli have at least 3 times higher amylase activity than the donor strain, of B. stearothermophilus. About 75％ of the $\alpha$-amylase produced by the constructed strain of E. coli was localized in the periplasm and it was found that the enzymes can be released by an osmotic shock using EDTA. The enzymatic properties of L-amylase produced in E. coli were very similar to those produced by B. stearothermophilus in terms of optimum temperature, heat stability and molecular weight.

• Korean Journal of Plant Tissue Culture
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• v.27 no.3
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• pp.219-226
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• 2000

Lycopersicon peruvianum has a gametophytic self-incompatibility (GSI) mechanism controlled by a single genetic locus (S locus) with multiple alleles. S RNases, an allelic series of abundant stylar proteins, are products of the S locus in L. peruvianum and other Solanaceous plants. The $S_{11}$ RNase gene from L. peruvianum was introduced into a self-compatible (SC) species (Lycopersicon esculentum) to examine whether the expression pattern in the heterologous host mimics that in L. peruvianum. The resultant transgenic L. esculentum plants expressed the introduced gene highly in their styles, which is similar manner to the expresion in L. peruvianum. The $S_{11}$ RNase gene was expressed in the syle at a similar stage of flower development in both transgenic plants of L. esculentum and L. peruvianum without any morphological changes.