• Journal of Microbiology and Biotechnology
• /
• 제6권1호
• /
• pp.36-42
• /
• 1996

The production of sorbitol by permeabilized cells of Zymomonas mobilis immobilized in $\kappa$-carrageenan was investigated. Cetyltrimethylammoniumbromide (CTAB) permeabilized cells were treated with glutaraldehyde prior to immobilization for cross-linking of enzymes, glucose-fructose oxidoreductase (GFOR) in cells. Rigidity of the immobilized beads was increased two-fold with 90$\%$ conversion efficiency by the additions of 40$\%$ (w/v) polyols (glycerol 25 g + propylene glycol 15 g) to 60$\%$ (w/v) distilled water containing 2.5$\%$ (w/v) $\kappa$-carrageenan as a final concentration, prior to immobilization. $\kappa$-Carrageenan beads entrapping permeabilized cells were dried to improve bead rigidity and storage stability. During s6mi-batch process for 72 h with dry beads, there was an improvement of the loss of enzyme activity (less than 10$\%$). In batch process, the kinetic results of $K_m.fructose$ value for the free cells, wet beads and dry $\kappa$-carrageenan beads were 71.7, 72.4 and 116.7 g/l, respectively. Higher productivity was obtained with two-stage continuous packed bed reactors with both wet and dry $\kappa$-carrageenan beads at 25.00 and 21.15 g/l/h, respectively, when measured at second stage.

• Journal of Microbiology and Biotechnology
• /
• 제26권7호
• /
• pp.1267-1277
• /
• 2016

Currently, enzymatic synthesis of lactulose, a synthetic prebiotic disaccharide, is commonly performed with glycosyl hydrolases. In this work, a new type of lactulose-producing biocatalyst was developed by displaying β-galactosidase from Bacillus stearothermophilus IAM11001 (Bs-β-Gal) on the surface of Bacillus subtilis 168 spores. Localization of β-Gal on the spore surface as a fusion to CotX was verified by western blot analysis, immunofluorescence microscopy, and flow cytometry. The optimum pH and temperature for the resulting spore-displayed β-Gal was 6.0 and 75℃, respectively. Under optimal conditions, it showed maximum activity of 0.42 U/mg spores (dry weight). Moreover, the spore-displayed CotX-β-Gal was employed as a whole cell biocatalyst to produce lactulose, yielding 8.8 g/l from 200 g/l lactose and 100 g/l fructose. Reusability tests showed that the spore-displayed CotX-β-Gal retained around 30.3% of its initial activity after eight successive conversion cycles. These results suggest that the CotX-mediated spore-displayed β-Gal may provide a promising strategy for lactulose production.

• 한국균학회지
• /
• 제17권1호
• /
• pp.31-34
• /
• 1989

인삼 saponin중 조성비율이 가장 큰 ginsenoside Rb1만을 약효면에서 보다 우수한 ginsenoside Rd로 선택적 전환시킬 수 있는 Rhizopus japonicus를 대상으로 해서 인삼 saponin 전환효소의 최적생산조건을 검토하였다. Wheat bran 배지를 기본으로 할 경우 배양 5일에 가장 많은 효소가 생산되었으며, 인삼류의 천연유기물중에는 홍삼분말을, 저급당류중에는 xylose를, 다당류중에는 laminarin을 flavonoids중에는 naringin을, 질소원중에는 beef extract와 $KNO_3$를 첨가할 경우 인삼 saponin 전환효소가 가장 많이 생산되었다.

• Korean Chemical Engineering Research
• /
• 제54권6호
• /
• pp.817-821
• /
• 2016

Levodopa or L-3,4-dihydroxyphenylalanine (L-DOPA) is the direct precursor of the neurotransmitter dopamine. L-DOPA is a well-known neuroprotective agent for the treatment of Parkinson's disease symptoms. L-DOPA was synthesized using the enzyme, tyrosinase, as a biocatalyst for the conversion of L-tyrosine to L-DOPA and an electrochemical method for reducing L-DOPAquinone, the product resulting from enzymatic synthesis, to L-DOPA. In this study, three electrode systems were used: A glassy carbon electrode (GCE) as working electrode, a platinum, and a Ag/AgCl electrode as auxiliary and reference electrodes, respectively. GCE has been modified using electropolymerization of pyrrole to facilitate the electron transfer process and immobilize tyrosinase. Optimum conditions for the electropolymerization modified electrode were a temperature of $30^{\circ}C$ and a pH of 7 producing L-DOPA concentration 0.315 mM. After 40 days, the relative activity of an enzyme for electropolymerization remained 38.6%, respectively.

• KSBB Journal
• /
• 제28권5호
• /
• pp.282-286
• /
• 2013

The seaweed, Gelidium amansii, was fermented to produce bioethanol. Optimal pretreatment condition was determined as 94 mM $H_2SO_4$ and 8% (w/v) seaweed slurry at $121^{\circ}C$ for 60 min. The mono sugars of 40.4 g/L with 67% of conversion from total carbohydrate of 60.6 g/L with 80 g dw/L G. amansii slurry were obtained by thermal acid hydrolysis pretreatment and enzymatic saccharification. G. amansii hydrolysate was used as the substrate for ethanol production by Kluyveromyces marxianus KCTC 7150 and Candida tropicalis KCTC 7212 using 5L fermentor. The ethanol productions by K. marxianus KCTC 7150 and C. tropicalis KCTC 7212 were 17.8 g/L with $Y_{EtOH}$ of 0.48 at 120 h and 19.3 g/L with $Y_{EtOH}$ of 0.50 at 120 h, respectively.

• Bulletin of the Korean Chemical Society
• /
• 제32권2호
• /
• pp.455-458
• /
• 2011

In previous report, with the aid of receptor-oriented pharmacophore-based in silico screening, we characterized three novel hPNMT inhibitors (YPN010, YPN016, and YPN017) and proposed that the hydrogen bonding interaction between inhibitors and side chain of Lys57 is very important to inhibitory activity of hPNMT. To confirm the importance of Lys57, mutant with substitution of Lys57 with Ala was cloned and binding study was performed for a K57A mutant of hPNMT using STD-NMR and fluorescence experiments. The binding constants for three novel inhibitors with mutant hPNMT were dramatically decreased compared to those with wild-type protein. K57A mutant-induced conversion of noradrenaline to adrenaline was suppressed about 95 % compared to wild-type hPNMT. Mutagenic analysis using a K57A mutant confirmed the importance of the Lys57 residue in binding of the inhibitor candidate to hPNMT as well as enzymatic activity of hPNMT, implying that these results are consistent with our binding model.

• KSBB Journal
• /
• 제29권4호
• /
• pp.297-302
• /
• 2014

The autoclaving assisted by an irradiation pretreatment method was developed without toxic chemicals to produce fermentable sugars for their conversion to bioethanol. In the first step, electron beam irradiation (EBI) of rice straw was performed at various doses. The electron beam-irradiated rice straw was then autoclaved with DI water at $120^{\circ}C$ for 1 h. A total sugar yield of 81% was obtained from 300 kGy electron beam-irradiated rice straw after 72 h of enzymatic hydrolysis by Cellulase 1.5L (70 FPU/mL) and Novozyme-188 (40 CbU/mL). Also, the removal of hemicellulose and lignin was 32.0% and 32.5%, respectively. This result indicates that the environmentally-friendly pretreatment method of rice straw by an electron beam irradiation could be applied for bioethanol production in plant.

• Mass Spectrometry Letters
• /
• 제1권1호
• /
• pp.29-32
• /
• 2010

Cross reacting antibodies can cause an overestimation of the results of immunoassays. Therefore, alternative methods are needed for the accurate quantification of steroids. Gas chromatography combined with isotope-dilution mass spectrometry (GC-IDMS) is developed to quantify urinary active androgens, testosterone, epitestosterone and dihydrotestosterone, which are clinically relevant androgens to both hair-loss and prostate diseases. The method devised involves enzymatic hydrolysis with $\beta$-glucuronidase, solid-phase extraction, liquid-liquid extraction using methyl tert-butyl ether and subsequent conversion to pentafluorophenyldimethylsilyl-trimethylsilyl (flophemesyl-TMS) derivatives for sensitive and selective analysis in selected-ion monitoring mode. Flophemesyl-TMS derivatization not only eliminates matrix interference but also has a good peak resolution within a 6 min-run. A selective and sensitive GC technique with flophemesyl-TMS derivatives also allows accurate quantitative analysis of three active androgens when combined with IDMS. The limit of quantification of the three analytes was <50 pg/mL, and extraction recoveries ranged from 91.9 to 102.1%. The precision and accuracy were 1.2~6.5% and 89.0~106.7%, respectively. This GC-IDMS method can be useful for evaluating the drug efficacy and monitoring the biological processes responsible for male-pattern baldness and prostate diseases.

• 한국미생물·생명공학회지
• /
• 제19권5호
• /
• pp.514-520
• /
• 1991

Extrusion시켜 구조변형시킨 전분(질)을 기질로 cyclodextrin glucanotransferse(CGTase)를 활용하여 액화과정을 거치지 않고 직접 cyclodextrin을 합성하는 불용성 extrusion된 전분-수용성 CGTase로 구성된 불균일상 효소반응계에 관하여 연구하였다. Extrusion된 전분을 기질로 이용할 경우 기존의 액화전분을 기질로하는 균일상 효소반응계에서 보다 현저히 증가된 CD 농도, 수율 그리고 합성속도를 얻을 수 있었으며, extrusion된 전분 기질농도가 100g/l일 때 CD생성량과 수율은 각각 54g/l와 0.54였다.

• Journal of Microbiology and Biotechnology
• /
• 제18권7호
• /
• pp.1245-1251
• /
• 2008

Oxalate decarboxylases (OXDCs) (E.C. 4.1.1.2) are enzymes catalyzing the conversion of oxalate to formate and $CO_2$. The OXDCs found in fungi and bacteria belong to a functionally diverse protein superfamily known as the cupins. Fungi-originated OXDCs are secretory enzymes. However, most bacterial OXDCs are localized in the cytosol, and may be involved in energy metabolism. In Agrobacterium tumefaciens C58, a locus for a putative oxalate decarboxylase is present. In the study reported here, an enzyme was overexpressed in Escherichia coli and showed oxalate decarboxylase activity. Computational analysis revealed the A. tumefaciens C58 OXDC contains a signal peptide mediating translocation of the enzyme into the periplasm that was supported by expression of signal-peptideless and full-length versions of the enzyme in A. tumefaciens C58. Further site-directed mutagenesis experiment demonstrated that the A. tumefaciens C58 OXDC is most likely translocated by a twin-arginine translocation (TAT) system.