• Microbiology and Biotechnology Letters
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• v.45 no.3
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• pp.226-235
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• 2017

Carotenoids such as phytoene, lycopene, and ${\beta}-carotene$ are used as food colorants, animal feed supplements, and for human nutrition and cosmetic purposes. Previously, we reported the isolation of a novel marine bacterium, Kocuria gwangalliensis, which produces a pink-orange pigment. Phytoene desaturase (CrtI), encoded by the gene crtI, catalyzes lycopene formation from phytoene and is an essential enzyme in the early steps of carotenoid biosynthesis. CrtI is one of the key enzymes regulating carotenoid biosynthesis and has been implicated as a rate-limiting enzyme of the pathway in various carotenoid synthesizing organisms. Here, we report the cloning of the crtI gene responsible for lycopene biosynthesis from K. gwangalliensis. The gene consisted of 1,584 bases encoding 527 amino acid residues. The nucleotide sequence of the crtI gene was compared with that of other species, including Kocuria rhizophila and Myxococcus xanthus, and was found to be well conserved during evolution. An expression plasmid containing the crtI gene was constructed (pCcrt1), and Escherichia coli cells were transformed with this plasmid to produce a recombinant protein of approximately 57 kDa, corresponding to the molecular weight of phytoene desaturase. Lycopene biosynthesis was confirmed when the plasmid pCcrtI was co-transformed into E. coli containing the plasmid pRScrtEB carrying the crtE and crtB genes required for lycopene biosynthesis. The results from this study will provide valuable information on the primary structure of K. gwangalliensis CrtI at the molecular level.

• Food Science and Preservation
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• v.18 no.2
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• pp.184-190
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• 2011

Limonoid UDP-glucosyltransferase (LUGT) is an enzyme that converts limonoids into their corresponding glucosides and ultimately ameliorates limonoid bitterness in Citrus species. In this paper, the LUGT gene was cloned via PCR from 10 Jeju Citrus species. All the deduced glucosyltransferase proteins harbored a highly conserved plant secondary product glucosyltransferase (PSPG) motif within the C terminal region. Phylogenetic analysis based on the amino acid sequence comparison of the LUGT proteins from 10 Citrus species generated three distinct types. The expression patterns of LUGT gene in three representative species from each type were quite different with that of C. unshiu Marc. cv. Miyagawawase(Gungcheon), which his without distinctive juice delayed bitterness. Ourresultssho wth at some Citrus speciessuchas Citrusleiocarpa HORT(Bingul), Citruserythrosa HORT (Dongjunggul), and Citrustachibana TANAKA(Honggul) end emicin Jeju maybe susceptible to intense juice delayed bitterness due to delay inexpression of LUGT.

• Korean Journal of Medicinal Crop Science
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• v.20 no.5
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• pp.359-364
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• 2012

This study were carried out to find bolting response of cultivation in different regions and to isolate FLC (FLOWERING LOCUS C) homologs in Angelica gigas Nakai. The mean temperature of different regions, ordering in altitude, were as follows: 100 m > 350 m > 530 m > 700 m. The largest amount of rainfall was occurred in the region of 350 m while the longest time of sunshine was occurred in the region of 100 m. The content of soil chemical properties in regions showed pH 6.2 ~ 7.4, T-N 0.17 ~ 26, organic mater $1{\sim}32gkg^{-1}$, $P_2O_5$ ${151{\sim}664_{mgkg}}^{-1}$, exchangeable potassium and calcium and magnesium were 0.78 ~ 1.15, 3.9 ~ 10.0, ${0.7{\sim}3.2_{cmol}}^{+kg-1}$. L5 line of A. gigas was occurred in bolting at all regions, but the bolting ratio was 60.0% in 700 m region with non-mulching treatment. Manchu of A. gigas was not occurred in bolting at all regions. The accumulation bolting ratio of L5 line by non-mulching was higher than that of mulching as 90.4% and 72.8% in 100 m region. The MADS-box transcription factor FLC is one of the well-known examples as a strong floral repressor. We decided to isolate FLC homologs from A. gigas as a starting point of flowering mechanism research of this plant. We have isolated two RT-PCR products which showed very high amino acid sequence homology to Arabidopsis FLC.

• Korean Journal of Clinical Pharmacy
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• v.20 no.3
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• pp.235-241
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• 2010

Bambuterol hydrochloride, dimethylcarbamic acid 5-[2-(1,1-dimethylethyl)amino-1-hydroxyethyl]-1,3-phenylene ester hydrochloride, is the prodrug of active ${\beta}_2$-adrenergic metabolite terbutaline. The purpose of the present study was to evaluate the bioequivalence of two bambuterol hydrochloride tablets, $Bambec^{(R)}$ tablet 10 mg (Yuhan Co., Ltd.) and Bambucol tablet 10 mg (Sam Chun Dang Pharm. Co., Ltd.), according to the guidelines of Korea Food and Drug Administration (KFDA). In vitro release of bambuterol from two bambuterol hydrochloride formulations was tested using KP VIII Apparatus II method with various dissolution media. Twenty eight healthy male Korean volunteers, $23.86{\pm}1.65$ years in age and $68.98{\pm}9.58$ kg in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After two tablets containing 10 mg as bambuterol hydrochloride were orally administered, blood samples were taken at predetermined time intervals, and the concentrations of bambuterol in serum were determined using column switching HPLC with UV detector. The dissolution profiles of two formulations were similar in all tested dissolution media. The pharmacokinetic parameters such as $AUC_t$, $C_{max}$ and $T_{max}$ were calculated, and ANOVA test with K-BE Test 2002 was utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_t$, $C_{max}$ and untransformed $T_{max}$. The results showed that the differences between two formulations based on the reference drug, $Bambec^{(R)}$, were -8.10%, -3.82% and 12.65% for $AUC_t$, $C_{max}$ and $T_{max}$, respectively. There were no sequence effects between two formulations in these parameters. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log 0.8 to log 1.25 (i.e., log 0.8093~log 1.0302 and log 0.8564~log 1.1280 for $AUC_t$ and $C_{max}$, respectively). Thus, the criteria of the KFDA bioequivalence guideline were satisfied, indicating Bambucol tablet 10 mg was bioequivalent to $Bambec^{(R)}$ tablet 10 mg.

• The Plant Pathology Journal
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• v.36 no.4
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• pp.323-334
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• 2020

Lysin motif (LysM) proteins are reported to be necessary for the virulence and immune response suppression in many herbaceous plant pathogens, while far less is documented in woody plant pathogens. In this study, we preliminarily characterized the molecular function of a LysM protein LtLysM1 in woody plant pathogen Lasiodiplodia theobromae. Transcriptional profiles revealed that LtLysM1 is highly expressed at infectious stages, especially at 36 and 48 hours post inoculation. Amino acid sequence analyses revealed that LtLysM1 was a putative glycoprotein with 10 predicted N-glycosylation sites and one LysM domain. Pathogenicity tests showed that overexpressed transformants of LtLysM1 displayed increased virulence on grapevine shoots in comparison with that of wild type CSS-01s, and RNAi transformants of LtLysM1 exhibited significantly decreased lesion length when compared with that of wild type CSS-01s. Moreover, LtLysM1 was confirmed to be a secreted protein by a yeast signal peptide trap assay. Transient expression in Nicotiana benthamiana together with protein immunoblotting confirmed that LtLysM1 was an N-glycosylated protein. In contrast to previously reported LysM protein Slp1 and OsCEBiP, LtLysM1 molecule did not interact with itself based on yeast two hybrid and co-immunoprecipitation assays. These results indicate that LtLysM1 is a secreted protein and functions as a critical virulence factor during the disease symptom development in woody plants.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.7
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• pp.1054-1062
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• 1999

The effects of dry roasting whole faba beans (WFB) and whole lupin seeds (WLS) at 110, 130 or $150{^{\circ}C}$ for 15, 30 or 45 min on rumen (RDCP%), estimated intestinal (IDCP%) and total tract disappearance of CP (TDCP%) and intestinal availability (IARUCP%) of rumen undegraded CP (RUCP%) were determined. The RDCP values were estimated by in sacco technique by incubating nylon bags for 8, 12 and 24 h in the rumen of dairy cows. The IDCP and IARUCP values were estimated using a sequence of ruminal incubation, in vitro incubation in acid-pepsin for 1 h and then in pancreatin for 24 h of three-step in vitro procedure technique. Dry roasting at 130 and $150^{\circ}C$ decreased RDCP with correspondingly increasing IDCP. The IDCP value generally increased from 12.3(raw) to 8.6, 14.8 and 39.6% (WFB) and from 28.3 (raw) to 33.7, 36.2 and 56.2% (WLS) at 8 h rumen incubation; from 2.9 (raw) to 2.9, 4.6 and 23.3% (WFB) and from 19.6 (raw) to 19.0, 24.0 and 46.6% (WLS) at 12 h rumen incubation; from 1.3 (raw) to 1.9, 1.7 and 11.0% (WFB) and from 4.4 (raw) to 4.2, 10.7 and 36.7% (WLS) at 12 h rumen incubation as the temperatures rose to 110, 130 and $150{^{\circ}C}$ respectively. The TDCP values were always high and increased by time in the rumen, the average values of which were 97.9, 96.6; 99.2, 96.9 and 99.6, 98.7% for WFB and WLS, respectively, at 8, 12 and 24 h rumen incubation. But within the same retention time, TDCP was generally unchanged. The average IARUCP increased from 87.3 (raw) to 87.4, 88.7 and 92.0% (WFB); from 87.6 (raw) to 88.9, 91.5 and 93.0% (WLS) at roasting temperatures of 110, 130 and $150{^{\circ}C}$, respectively. It was concluded that dry roasting can shift the digestion of CP from rumen to the lower gastrointestinal tract without depressing the digestion of RUCP. The best processing condition in this study was dry roasting at $150{^{\circ}C}$ for 45 min in terms of effects on the disappearances and availability of CP. Research data on intestinal availability of individual amino acids need to be further investigated.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.3
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• pp.198-204
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• 2000

In order to utilize by-products which would normally be discarded in marine processing plants, cod teiset protein was hydrolyzed and antioxidative actiTity of the hydrolysate was investigated. AntioxidatiTe peptide was isolated using ultrafiltration membrane, ion-exchange chromatography on a SP-Sephadex C-25 column, gel filtration on a Sephadex G-15 column, high performance liquid chromatography on an ODS column, and capillary electrophoresis chromatography. Antioxidative activities of the cod teiset hydrolysate were compared with ${\alpha}-tocopherol$, one of the commercial antioxidant. The hydrolysate passed through a membrane with molecular weight cut-off (MWCO) 1 kDa was shown the strongest antioxidative activity, and the activity was higher $10{\%}$ as compared with ${\alpha}-tocopherol$. In addition, the peptide isolated by ion-exchange chromatography, gel filtration, and HPLC, respectively, was higher $53{\%}$ as compared with ${\alpha}-tocopherol$, and the amino acid sequence was Ser-Asn-Pro-Glu-Trp-Ser-Trp-Asn.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.11
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• pp.1544-1550
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• 2008

The retinoids (vitamin A and its derivatives) play a critical role in vision, growth, reproduction, cell differentiation and embryonic development. Using the IMpRH panel, porcine cellular retinol binding protein genes 5 and 7 (RBP5 and RBP7) were assigned to porcine chromosomes 5 and 6, respectively. The complete coding sequences (CDS) of the RBP5 and RBP7 genes were amplified using the reverse transcriptase polymerase chain reaction (RT-PCR) method, and the deduced amino acid sequences of both genes were compared to human corresponding proteins. The mRNA distributions of the two genes in adult Wuzhishan pig tissues (lung, skeletal muscle, spleen, heart, stomach, large intestine, lymph node, small intestine, liver, brain, kidney and fat) were examined. A total of nine single nucleotide polymorphisms (SNPs) were identified in two genes. Three of these SNPs were analyzed using the polymerase chain reaction-restriction-fragment length polymorphism (PCR-RFLP) method in Laiwu, Wuzhishan, Guizhou, Bama, Tongcheng, Yorkshire and Landrace pig breeds. Association analysis of genotypes of these SNP loci with economic traits was done in our experimental populations. Significant associations of different genotypes of $RBP5-A/G^{63}$, $RBP5-A/G^{517}$ and $RPB5-T/C^{intron1-90}$ loci with traits including maximum carcass length (LM), minimum carcass length (LN), marbling score (MS), back fat thickness at shoulder (SBF), meat color score (MCS) and hematocrit (HCT) were detected. These SNPs may be useful as genetic markers in genetic improvement for porcine production.

• Korean Journal of Microbiology
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• v.43 no.2
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• pp.83-90
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• 2007

Sphingomonas sp. KS 301, which was isolated from oil contaminated soil, was shown to have five different SODs (SODI, II, III, IV, V) which can be separated by DEAE-Sepharose chromatography, and SOD III was finally purified in this study by ammonium sulfate precipitation, DEAE-Sepharose chromatography, Superose 12 gel filtration and Uno-Q1 ion exchange chromatography. The molecular weight of SOD III was 23 kDa as determined by SDS-PAGE and the apparent molecular weight of the native enzyme was estimated to be approximately 71 kDa by Superose-12 gel filtration chromatography. These data suggest that the purified SOD consists of at least two subunits. The specific activity of the SOD III was higher than Mn type or Fe type SOD of Escherichia coli by 5 fold. To determine the type of SOD III, inhibitory effects of $NaN_{3},\;H_{2}O_{2},\;KCN$ were examined. 10 mM $NaN_{3}$ was able to inhibit 56% of the SOD III activity, which indicates that this SOD is Mn type. The optimum pH of the SOD III was 7.0 and the optimum temperature was $20^{\circ}C$. N-terminal amino acid sequence of purified SOD III was most similar to those of Psudomonase ovalis and Vibrio cholerae among bacteria.

• Journal of Microbiology and Biotechnology
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• v.19 no.11
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• pp.1295-1300
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• 2009

A 2,172-bp full-length gene (aga-F78), encoding a protease-resistant $\alpha$-galactosidase, was cloned from Rhizopus sp. F78 and expressed in Escherichia coli. The deduced amino acid sequence shared highest identity (45.0%) with an $\alpha$-galactosidase of glycoside hydrolase family 36 from Absidia corymbifera. After one-step purification with a Ni-NTA chelating column, the recombinant Aga-F78 migrated as a single band of ~82 and ~210 kDa on SDS-PAGE and nondenaturing gradient PAGE, respectively, indicating that the native structure of the recombinant Aga-F78 was a trimer. Exhibiting the similar properties as the authentic protein, purified recombinant Aga-F78 was optimally active at $50^{\circ}C$ and pH 4.8, highly pH stable over the pH range 5.0-10.0, more resistant to some cations and proteases, and had wide substrate specificity (pNPG, melidiose, raffinose, and stachyose). The recombinant enzyme also showed good hydrolytic ability to soybean meal, releasing galactose of $415.58\;{\mu}g/g$ soybean meal. When combined with trypsin, the enzyme retained over 90% degradability to soybean meal. These favorable properties make Aga-F78 a potential candidate for applications in the food and feed industries.