• Biotechnology and Bioprocess Engineering:BBE
• /
• v.11 no.5
• /
• pp.442-448
• /
• 2006

Human lactoferrin (hLf) is an iron-binding glycoprotein that has been considered to play many biological roles in the human, including the stimulation of the immune system, antimicrobial and anti-inflammatory effects, and regulation of iron absorption. We generated transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing a functional hLf protein using the signal peptide sequence from the endoplasmic reticulum and driven by an oxidative stress-inducible SWPA2 promoter which is highly expressed in plant cell cultures. The production of hLf increased proportionally to cell growth and showed a maximal level (up to 3.6% of total soluble protein) at the stationary phase in suspension cultures. Full-length hLf protein was identified by immunoblot analysis in transgenic cell cultures of Siberian ginseng. Recombinant hLf (rhLf) was purified from suspension cells of Siberian ginseng by ammonium sulfate precipitation, cation-exchange and gel filtration chromatography. N-terminal sequences of rhLf were identical to native hLf (nhLf). The overall monosaccharide composition of rhLf showed the presence of plant specific xylose while sialic acid is absent. Antibacterial activity of purified rhLf was higher than that of nhLf. Taken together, we anticipate that medicinal Siberian ginseng cultured cells, as demonstrated by this study, will be a biotechnologically useful source for commercial production of functional hLf not requiring further purification.

• Fisheries and Aquatic Sciences
• /
• v.1 no.2
• /
• pp.201-208
• /
• 1998

A protease, which had no tryptic and chymotryptic activity, was purified from the hepatopancreas of shrimp, P. japonicus, through ammonium sulfate fractionation, Q­Sepharose ionic exchange, benzamidine Sepharose 6B affinity, and Sephacryl S-100 gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 24 kDa by gel filtration and showed a single peptide band by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The protease had a low ratio of acidic to basic amino acids, which is different with pro teases from marine animals. The enzyme was partially inhibited by benzamidine, tosyl-L-lysine chioromethyl ketone (TLCK), phenylmethylsulfonyl fluoride (PMSF), soybean trypsin inhibitor (SBTI), and pepstatin. The enzyme did not have any activity against benzoyl-D,L-arginine p-nitroanilide (BAPNA) or benzoyl-L-tyrosine ethyl ester (BTEE) which is a specific substrate of trypsin and chymotrypsin, respectively. However, the enzyme showed activity forward N-CBZ-L-tyrosine p-nitrophenyl ester (CBZ-Tyr-pNE), N­CBZ-L-tryptophan p-nitrophenyl ester (CBZ-Trp-pNE), and N-CBZ-L-proline p-nitrophenyl ester (CBZ-Pro-pNE). The protease did not showed tryptic and chymotryptic activity, which was not reported in shrimp hepatopancreas.

• KSBB Journal
• /
• v.10 no.3
• /
• pp.271-282
• /
• 1995

The authentic hGH converted from met-hGH by immobilized ApM was purified by successive chromatographic processes based on the differences in isoelectric points, hydrophobicities and charges. The final recovery yield was about 14.1% and the specific activity of the purified hGH was 2.75IU per mg when assayed by enzyme immunoassay. The purified hGH was verified to be authentic hGH through the analysis of amino acid composition, amino-terminal amino acid sequence, carboxy-terminal amino acid and tryptic peptide map. The purity of purified hGH was higher than that of commercial hGH when assessed by SDS-PAGE, PAGE, IEF and HSGF. In weight-gain assay and tibia test with hypophysectomized rats, the hGH produced in this study showed the same growth effect as the commercial hGH.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.22 no.2
• /
• pp.226-233
• /
• 1993

Enzymatic hydrolysates of food proteins (defatted soybean cake, egg albumin and casein) were tested for inhibitory activity against angiotensin-I converting enzyme (ACE). Food proteins were hydrolysed with complex enzyme, bromelain, alcalase, $\alpha$-chymotrypsin, trypsin, papain and pepsin by heating method. The hydrolysates obtained from the treatment of complex enzyme and bromelain showed the higher ACE inhibitory activity. ACE inhibitory activity of hydrolysates exhibited a tendency to be increased until 8hrs and increased with increment of concentration. The activity was also stable by heat treatment at 10$0^{\circ}C$ for 20min. Molecular weight of active fraction was about 1, 400 and defatted soybean cake hydrolysate below 1, 400 in case of defatted soybean cake hydrolysate treated with alcalase. Amino acid of the active fractions was abundant in Asp, Glu, Lys, lle, Leu, Ala and Val.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.33 no.4
• /
• pp.239-248
• /
• 2019

There are no published data on Gyeongokgo (GOK) safety or efficacy despite being commonly use. The Gyeongokgo (GOK) is commonly used in traditional Korean medicine to promote a health qi and blood, but their objective data was not sufficient in clinical field. To investigate the safety and efficacy of GOK with hematologic and serologic testing and the change of the quality of life in healthy individuals. Randomized, subject-assessor-blind, placebo-controlled, single-center pilot study Participants and Interventions 29 healthy volunteer subjects were randomly placed into the GOK group (n = 20) or placebo control group (n = 9) and instructed to take one treatment packet (GOK or placebo) twice daily for 4 weeks. Subjects were assessed using the Fatigue Severity Scale (FSS) and Short Form 36 Health Survey (SF-36) and underwent hematologic and serologic tests and body composition analysis. The FSS total score (p = 0.093) and SF-36 general health index (p = 0.002) were improved following treatment in the GOK group. Post-treatment thyroid-stimulating hormone levels were increased in the GOK group compared with pre-treatment levels (p = 0.0042). C-reactive protein levels decreased (p = 0.0256) in the GOK group compared with that the placebo group. In time-series tests, GOK did not affect post-prandial serum triglycerides, glucose, insulin, or C-peptide levels. Notably, elevations in serum fasting triglycerides at 2- (p = 0.0333) and 4-hours (p = 0.0414) post-prandial were lower than those in the placebo group. GOK reduced fatigue levels and did not significantly affect laboratory test results performed to measure safety, serum glucose, and lipid profiles. Post-meal triglyceride levels were effectively reduced with treatment.

• Animal Bioscience
• /
• v.36 no.7
• /
• pp.1130-1142
• /
• 2023

Objective: Cow urine possesses several bioactive properties but the responsible components behind these bioactivities are still far from identified. In our study, we tried to identify the possible components behind the antimicrobial activity of cow urine by exploring the peptidome and metabolome. Methods: We extracted peptides from the urine of Sahiwal cows belonging to three different physiological states viz heifer, lactation, and pregnant, each group consisting of 10 different animals. The peptides were extracted using the solid phase extraction technique followed by further extraction using ethyl acetate. The antimicrobial activity of the aqueous extract was evaluated against different pathogenic strains like Staphylococcus aureus, Escherichia coli, and Streptococcus agalactiae. The safety of urinary aqueous extract was evaluated by hemolysis and cytotoxicity assay on the BuMEC cell line. The urinary peptides were further fractionated using high-performance liquid chromatography (HPLC) to identify the fraction(s) containing the antimicrobial activity. The HPLC fractions and ethyl acetate extract were analyzed using nLC-MS/MS for the identification of the peptides and metabolites. Results: A total of three fractions were identified with antimicrobial activity, and nLC-MS/MS analysis of fractions resulted in the identification of 511 sequences. While 46 compounds were identified in the metabolite profiling of organic extract. The urinary aqueous extract showed significant activity against E. coli as compared to S. aureus and S. agalactiae and was relatively safe against mammalian cells. Conclusion: The antimicrobial activity of cow urine is a consequence of the feeding habit. The metabolites of plant origin with several bioactivities are eliminated through urine and are responsible for their antimicrobial nature. Secondly, the plethora of peptides generated from the activity of endogenous proteases on protein shed from different parts of tissues also find their way to urine. Some of these sequences possess antimicrobial activity due to their amino acid composition.

• Journal of Plant Biotechnology
• /
• v.1 no.1
• /
• pp.20-30
• /
• 1999

In order to modify lipid production of Perilla qualitatively as well as quantitatively by genetic engineering, genes involved in carbon metabolism were isolated and characterized. These include acyl-ACP thioesterases from Perilla frutescens and Iris sp., four different $\beta$-ketoacyl- ACP synthases from Perilla frutescens, and two $\Delta$15 a-cyl-ACP desaturases(Pffad7, pffad3). Δ15 acyl-ACP desa turase (Δ15-DES) is responsible for the conversion of linoleic acid (18:2) to $\alpha$-linolenic acid (ALA, 18:3). pffad 3 encodes Δ15 acyl-desaturase which is localized in ER membrane. On the other hand, Pffad7 encodes a 50 kD plastid protein (438 residues), which showed highest sequence similarity to Sesamum indicum fad7 protein. Northern blot analysis revealed that the Pffad7 is highly expressed in leaves but not in roots and seeds. And Pffad3 is expressed throughout the seed developmental stage except very early and fully mature stage. We constructed Pffad7 gene under 355 promoter and Pffad3 gene under seed specific vicillin promoter. Using Pffad7 construct, Perilla, an oil seed crop in Korea, was transformed by Agrobacterium leaf disc method. $\alpha$-linolenic acid contents increased in leaves but decreased in seeds of transgenic Perilla. Currently, we are transforming Perilla with Pffad3 construct to change Perilla seed oil composition. We isolated three ADP-glucose pyrophosphorylase (AGP) genes from Perilla immature seed specific cDNA library. Nucleotide sequence analysis showed that two of three AGP (Psagpl, Psagp2) genes encode AGP small subunit polypeptides and the remaining (Plagp) encodes an AGP large subunit. PSAGPs, AGP small subunit peptide, form active heterotetramers with potato AGP large subunit in E. coli expressing plant AGP genes.

• Journal of Plant Biotechnology
• /
• v.38 no.4
• /
• pp.263-271
• /
• 2011

Rice is one of the most important food crops since it is consumed by approximately 60% of the world's population. The most abundant component of rice grain is starch that is an important source of energy. The second abundant component is protein, which is an important protein source for people in many developing countries that rarely take animal protein. However, the rice protein lacks the essential amino acid lysine. Therefore, nutritional improvement in the essential amino acid composition of rice proteins is required. On the other side, rice grain has attracted attention as a diet and health food in developed countries, because its proteins have superior physiological and food processing properties. Thus, nutritional improvements in rice seed proteins by changing amino acid composition or introducing an useful protein or peptide have been studied. This review aims at assessing the current research status of biosynthesis, accumulation, genetic improvement of seed storage proteins by mutation or genetic engineering in rice.

• Journal of Life Science
• /
• v.19 no.11
• /
• pp.1672-1678
• /
• 2009

The purified antifungal substances produced by Bacillus amyloliquefaciens IUB158-03 was positive to ninhydrin but negative to aniline, suggesting that the antifungal substance could be a peptide. FAB-MS, UV adsorption spectrum, and amino acid composition analysis revealed that the molecular weight of the antifungal substance was 1042 and that maximal adsorption was at 220 nm and 277 nm. The antifungal substance was composed of $Asn_3$, $Gln_2$, $Ser_1$, $Gly_1$, and $Tyr_1$. The composition and structural characteristics of antifungal substance were analysed by $^1H$-NMR spectrum, $^1H$-COSY, HMQC, which revealed that the compound belongs to the iturin A family. Temperature and pH had little effect on the stability of the antifungal substance in the ranges of $-70{\sim}121^{\circ}C$ and pH 6.0~10.0, respectively. It showed strong antibiotic activity against fungi. An in vitro cytotoxicity test using NIH3T3 cell showed that the antifungal substance does not have cytotoxicity. The number of circulating leukocytes and the hematobiological analysis of the mice administered with the antifungal substances was similar to those of the control group, indicating no cytotoxicity in vivo. Therefore, the antifungal substances extracted from culture broth of Bacillus amyloliquefaciens IUB158-03 have future potential as biocontrol agents against plant diseases caused by fungi.

• Korean Journal of Food Science and Technology
• /
• v.46 no.5
• /
• pp.630-635
• /
• 2014

Pine nut oil (PNO) is well known to impart beneficial effects in overweight individuals, but the mechanisms underlying PNO-mediated weight loss remain unclear. To investigate how PNO promotes weight loss, its composition was determined by gas chromatography coupled with mass spectrometry (GC-MS). In addition, the effects of PNO on cytotoxicity, lipid accumulation, expression of lipid metabolism-related biomarkers, and leptin secretion were assessed in 3T3-L1 cells. GC-MS analyses revealed that PNO contains several components, including linoleic acid, oleic acid, palmitic acid, and stearic acid. Moreover, PNO did not have a cytotoxic effect on 3T3-L1 cells. However, it inhibited the expression of peroxisome proliferator-activated receptor (PPAR) and adipocyte protein 2 (aP2). Finally, PNO significantly increased leptin secretion in a dose-dependent manner. Taken together, these results support the notion that PNO is useful for weight management in overweight individuals.