• Journal of Applied Biological Chemistry
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• v.59 no.3
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• pp.173-178
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• 2016

Phenyllactic acid (PLA), which is a known antimicrobial compound, can be synthesized through the reduction of phenylpyruvic acid (PPA) by lactate dehydrogenase of lactic acid bacteria (LAB). PLA-producing LAB was isolated from coffee beans, and the isolated LAB was identified as Lactobacillus zeae Y44 by 16S rRNA gene sequence analysis. Cell-free supernatant (CFS) from L. zeae Y44 was assessed for both its capability to produce the antimicrobial compound PLA and its antifungal activity against three fungal pathogens (Rhizoctonia solani, Botrytis cinerea, and Colletotrichum aculatum). PLA concentration was found to be 4.21 mM in CFS when L. zeae Y44 was grown in MRS broth containing 5 mM PPA for 12 h. PLA production could be promoted by the supplementation with PPA and phenylalanine (Phe) in the MRS broth, but not affected by 4-hydroxy-phenylpyruvic acid, and inhibited by tyrosine as precursors. Antifungal activity assessment demonstrated that all fungal pathogens were sensitive to 5 % CFS (v/v) of L. zeae Y44 with average growth inhibitions ranging from 27.8 to 50.0 % (p<0.005), in which R. solani was the most sensitive with an inhibition of 50.0 %, followed by B. cinerea and C. aculatum. However, pH modification of CFS to pH 6.5 caused an extreme reduction in their antifungal activity. These results may indicate that the antifungal activity of CFS was caused by acidic compounds like PLA or organic acids rather than proteins or peptides molecules.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.2
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• pp.155-161
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• 2005

The inherent instability of metabolite production in plant cell culture-based bioprocessing is a major problem hindering its commercialization. To understand the extent and causes of this instability, this study was aimed at understanding the variability of anthocyanin accumulation during long-term subcultures, as well as within subculture batches, in Vitis vinifera cell cultures. Therefore, four cell line suspensions of Vitis vinifera L. var. Gamay Freaux, A, B, C and D, originated from the same callus by cell-aggregate cloning, were established with starting anthocyanin contents of $2.73\;\pm\;0.15,\;1.45\;\pm\;0.04,\;0.7\;\pm\;0.024\;and\;0.27\;\pm\;0.04$CV (Color Value)/g-FCW (fresh cell weight), respectively. During weekly subculturing of 33 batches over 8 months, the anthocyanin biosynthetic capacity was gradually lost at various rates, for all four cell lines, regardless of the significant difference in the starting anthocyanin content. Contrary to this general trend, a significant fluctuation in the anthocyanin content was observed, but with an irregular cyclic pattern. The variabilities in the anthocyanin content between the subcultures for the 33 batches, as represented by the variation coefficient (VC), were 58, 57, 54, and $84\%$ for V. vinifera cell lines A, B, C and D, respectively. Within one subculture, the VCs from 12 replicate flasks for each of 12 independent subcultures were averaged, and found to be $9.7\%$, ranging from 4 to $17\%$. High- and low-producing cell lines, VV05 and VV06, with 1.8-fold differences in their basal anthocyanin contents, exhibited different inducibilities to L-phenylalanine feeding, methyl jasmonate and light irradiation. The low-producing cell line showed greater potential in enhanced the anthocyanin production.

• Food Science of Animal Resources
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• v.41 no.4
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• pp.687-700
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• 2021

This study performed to evaluate the applicability of functional dairy food materials by comparing the calcium solubilization ability and anti-inflammatory effects of hydrolyzed casein protein. Commercial enzyme (Alcalase^®; Neutrase^®; Protamex^®; Flavourzyme^®) was added to the 10% casein solution to prepare the casein hydrolysates. Samples obtained every hour [1:200 (w/v)]. According to results of measuring the degree of hydrolysis (DH), all of four enzymatic hydrolysates increased rapidly from 30 to 40 min, and after 150 min, there were no change. Protamex^® and Neutrase^® had the highest DH compared to others enzymatic hydrolysates. After that, peptides obtained throughout a preparative liquid chromatography system. In the calcium solubility experiments, neutrase fraction (NF) 4 and NF7 showed similar activities with casein phosphopeptide (CPP). In vitro cell experiments showed that no cytotoxicity except for NF6. Also, the production of nitric oxide (NO) inhibited as the concentration of fraction samples increased. The cytokine (IL-1α, IL-6, and TNF-α) production was lower than lipopolysaccharide (+) group significantly. Therefore, the possibility of anti-inflammatory activity found in the hydrolyzed samples. According to the above experiments, NF3 and Protamex Fraction (PF) 3 selected. Amino acids selected throughout an AccQ-Tag system. As a result, 17 species of amino acids and several species of unknown amino acids identified. Both fractions had the highest content of phenylalanine. This study identified the potential of biologically active and functional peptides derived from casein that affect the food and dairy industry.

• Applied Biological Chemistry
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• v.43 no.4
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• pp.247-252
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• 2000

As functionality investigation of Korean traditional soybean fermentation foods, an antihypertensive peptide was separated during Chunggugjang fermentation by Bacillus subtilis CH-1023 and investigated inhibitory effect against angiotensin converting enzyme. After incubation at $20^{\circ}C,\;30^{\circ}C,\;40^{\circ}C,\;50^{\circ}C,\;60^{\circ}C$ for the $0{\sim}72$ hrs, protein content, protease activity and angiotensin converting enzyme inhibitory rate were determined. The protein content and protease activity were increased and reached maximum at 60 hrs fermentation with $40^{\circ}C$ and decreased after the 60 hrs fermentation. The optimum condition for antihypertensive peptide from Chunggugjang was appeared for 60 hrs at $40^{\circ}C$. Crude extract of Chunggugjang was partially purified by Amicon YM-3 membrane filtration and Sephadex G-10, G-25 gel filtration. The purified peptide showed inhibitory rate of 94.3% with 0.5 mg peptide content. The most prominent amino acid composition of the peptide from Chunggugjang was alanine, followed by phenylalanine, histidine.

• Journal of Korean Society of Forest Science
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• v.80 no.4
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• pp.436-444
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• 1991

To increase both the production of mushrooms and the usages of the residues (used media) for roughage, the pinchips(PC) and sawdusts(SD) of three Korean oak species were tested as cultivating media for Lentinus edodes with varing the composition of the media. The results are as follows. 1. Organic acids(tannic acid and citric acid), which were added in the PC medium to enrich the quality of mushrooms, did not increase the growths of the mycelia and the quality. 2. The mushrooms were able to be harvested from six months after beginning the cultivation. No significant differences were found among the qualities of the mushrooms produced by various treatments. 3. The contents of crude proteins(1.82-4.55%) in the used media were higher than those in woods (c.a. 0.7%), and the total digestible nutrients (44.0-46.0%) in the used medium were not much different from those in rice straws (c.a.48.0%). 4. The contents of some essential amino acids (methionine in the used medium of the tannic acid added PC or S.D. and isoleucine, phenylalanine and lysine in the used medium of the untreated PC or SD) were higher than those in the control (raw media). The contents of a few other essential amino acids (threonine, valine and arginine) were lower in the used media. Most nonessential amino acids in the used media, particularly in the used bed-log, appeared to decrease than those in the control.

• Journal of Plant Biotechnology
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• v.42 no.3
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• pp.135-153
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• 2015

The aromatic amino acids, which are composed of $\small{L}$-phenylalanine, $\small{L}$-tyrosine and $\small{L}$-tryptophan, are general components of protein synthesis as well as precursors for a wide range of secondary metabolites. These aromatic amino acids-derived compounds play important roles as ingredients of diverse phenolics including pigments and cell walls, and hormones like auxin and salicylic acid in plants. Moreover, they also serve as the natural products of alkaloids and glucosinolates, which have a high potential to promote human health and nutrition. The biosynthetic pathways of aromatic amino acids share a chorismate, the common intermediate, which is originated from shikimate pathway. Then, tryptophan is synthesized via anthranilate and the other phenylalanine and tyrosine are synthesized via prephenate, as intermediates. This review reports recent studies about all the enzymatic steps involved in aromatic amino acid biosynthetic pathways and their gene regulation on transcriptional/post-transcriptional levels. Furthermore, results of metabolic engineering are introduced as efforts to improve the production of the aromatic amino acids-derived secondary metabolites in plants.

• Korean Journal of Poultry Science
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• v.34 no.2
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• pp.119-128
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• 2007

The effects of microbial phytase on laying performance, egg quality, and ileal digestibility of nutrients and amino acids were examined at three levels of phytase (0, 300, 600 unit/kg) in 55-wk-old White Leghorn for 4 weeks. Egg productivity tended to increase with supplemental phytase compared to that of control. Daily feed intake of hens fed phytase also increased. Egg shell thickness was not significantly different among the treatments. Haugh unit and yolk color were not statistically different. However, egg shell breaking strength was high at phytase treatment. Excretion and absorption of nitrogen were no difference among all treatments, but those of phosphorus was higher in the phytase treatment than control. The digestibility was high at crude fiber, crude ash, calcium and phosphorus in nutrients, at lysine, methionine and phenylalanine in essential amino acids, and alanine, cystine, glutamic acid, glycine and tyrosine in non-essential amino acids. In conclusion, supplemental microbial phytase in laying hens diet may help to improve egg production and to decrease P of feces. But, further studies were needed to investigate on the digestibility.

• Korean Journal of Plant Tissue Culture
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• v.27 no.6
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• pp.453-459
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• 2000

Three rice (Oryza sativa L. var Dong-Jin) mutants (DTR1, DTR2, DTR3) resistant to S-methyltryptophan (5MT) were selected by mutagenized M3 seeds. The frequency of chlorophyll mutations induced by the EMS (0.2%) treatment performed 2 hours after flowering is clearly higher than that induced by other treatments in M1 generation. Progeny obtained from the self-pollinating of 5MT-resistant lines segregated with 3 : 1 of resistant to sensitive ratio. Furthermore, the ratio of homozygote to heterozygote in 5MT-resistant plants of the M4 generation was 1：2. These results show that 5MT resistance was inherited as a single dominant nuclear gene. The resistance was also expressed in callus derived from seeds. Total free amino acid content in homozygous seeds of DTR1 and DTR2 showed about 1.7 fold-increased compared to the wild-type seeds. In particular, the levels of phenylalanine and Iysine were, respectively, 6.2 and 3.2 times higher than those in the wild-type seeds. However, seeds of DTR3 had lower levels of free amino acid than the wild-type seeds. This result indicate that these mutants as a significant step towards the production of new rice with balanced amino acid content.

• Food Science and Industry
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• v.40 no.2
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• pp.83-89
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• 2007

저온발효와 고온발효에 의하여 만들어진 민속주를 증류하여 알콜돗를 45%로 같이 맞추어 성분함량을 비교하였다. 발효온도에 따른 환원당의 함량을 나타내고 있으며 저온발효 민속주는 15 mg/100 mL, 고온발효 민속주는 5 mg/100 mL로 나타났다. 즉 저온 발효에서 환원당의 양이 많았으며, 그러나 시중에 판매되고 있는 소주보다는 낮은 경향을 보였다. 향기 성분에 생성과정에 영향을 주는 Fusel alcohol을 볼 때 반적으로는 발효액에 아미노산이 없는 것보다는 많을 수록 많은 양의 fusel alcohol을 생성되기는 하지만, fusel alcohol을 생성하기위한 조건은 효모의 종류, 발효조건, 발효온도, 발효액의 조성 등의 복잡한 인자가 관련된다. 본 연구에 제조한 발효주를 소주의 아미노산과 비교할 때 아미노산 함량이 전반적으로 20-100도 낮은 함량치로 나타났다. 한약재 첨가로 인하여 면역증강, 살세포 반응 억제효과, 기능성 전통주로 좋은 발효주로 인정되지만 한약재특유의 강한 향기를 약화시키는 연구가 금후에 보완되어야 한다고 본다. 발효온도에 따른 아미노산의 함량은 저온발효 조미주에서 cysteine, valine, mrthionine, isoleucine, phenylalanine이 많았으며, 고온발효 조미주에서는 serine, glycine, leucine이 많았다. 한 alanine, tyrisine, lysine은 고온발효주에는 있으나, 저온발효주에서는 나타나지 않았다. 그러나 저온발효주와 고온 발효주 모두 시중에 판매되고 있는 소주보다는 아주 낮은 경향을 보였다. 저온발효와 고온발효에 의하여 만들어진 민속주를 증류하여 알콜 도수를 45%로 같이 맞추어 성분함량을 비교하였다. 발효온도에 따른 환원당의 함량을 나타내고 있으며 저온발효 민속주는 15 mg/100 mL, 고온발효 민속주는 5 mg/100 mL로 나타났다. 즉 저온 발효에서 환원당의 양이 많았으며, 그러나 시중에 판매되고 있는 소주보다는 낮은 경향을 보였다. 아미노산 실험에서는 소주와 45% 고온 발효주를 비교 할 때 serine, glycine, cysteine, methionine, phenylalanine 등은 100배, alanine, valine, isoleucine 80배, ammonia 20배 낮은 수치를 나타냈다. OD 측정의 실험결과 또한 control과 비교시 40% 발효주에서는 10배, 45% 고은 발효주에서는 100배 낮은 측정치가 나타났으므로 한약재의 색소성분을 휘발시키는 기술이 요구된다고 본다. 항균성실험에서 항균성 측정은 06 cm paper disk agar diffusion법을 이용하였으며, 43%의 발효주와, 45% 고온 발효주가 항균력이 가장 강력한 0.95 cm의 영향을 나타냈다. 사용한 사용한 Gram 양성, Gram 음성 균주는 Escherichia coli KCCM 11591를 제외하고는 0.8 - 0.95 cm로 항균력이 강했으며, Gram negitive의 Pseudomonas aeruginosa KCTC 1750 에서는 43% 발효주에는 0.95 cm, 45% 고은 발효주에는 0.95 cm의 항균성을 나타냈으며 관능평가에서도 가장 높게 났다. 관능평가에서는 45% 고온 발효주가 가장 높게 나타났으며, 항산화성 실험에 나타난 저온 45%의 갈색도의 측정과는 항산화성에서는 좀 다른 결과를 나타낸다. 그러나 항균성이 가장 높게 나타난 43-45%와 관능평가에서 가장 높게 나타난 45% 고온 발효주를 볼 때 본 연구에서는 고온 발효주 45%가 가장 우수한 전통주로 조사되었다. 발효주를 소주의 아미노산과 비교할 때 아미노산 함량이 전반적으로 100배 정도 낮은 함량치로 나타났으므로, 한약재 첨가로 인하여 면역증강, 살세포반응 억제효과, 기능성전통주로 좋은 발효주로 인정되지만 한약재특유의 강한 향기를 약화시켜서 부드럽고 은은한 전통발효주의 연구가 금후에 보완되어야 한다고 본다.

• Animal Bioscience
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• v.35 no.2
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• pp.224-235
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• 2022

Objective: Cage rearing has critical implications for the laying duck industry because it is convenient for feeding and management. However, caging stress is a type of chronic stress that induces maladaptation. Environmental stress responses have been extensively studied, but no detailed information is available about the comprehensive changes in plasma metabolites at different stages of caging stress in ducks. We designed this experiment to analyze the effects of caging stress on performance parameters and oxidative stress indexes in ducks. Methods: Liquid chromatography tandem mass spectrometry (LC/MS-MS) was used to determine the changes in metabolites in duck plasma at 5 (CR5), 10 (CR10), and 15 (CR15) days after cage rearing and traditional breeding (TB). The associated pathways of differentially altered metabolites were analyzed using Kyoto encyclopedia of genes and genomes (KEGG) database. Results: The results of this study indicate that caging stress decreased performance parameters, and the plasma total superoxide dismutase levels were increased in the CR10 group compared with the other groups. In addition, 1,431 metabolites were detected. Compared with the TB group, 134, 381, and 190 differentially produced metabolites were identified in the CR5, CR10, and CR15 groups, respectively. The results of principal component analysis (PCA) show that the selected components sufficiently distinguish the TB group and CR10 group. KEGG analysis results revealed that the differentially altered metabolites in duck plasma from the CR5 and TB groups were mainly associated with ovarian steroidogenesis, biosynthesis of unsaturated fatty acids, and phenylalanine metabolism. Conclusion: In this study, the production performance, blood indexes, number of metabolites and PCA were compared to determine effect of the caging stress stage on ducks. We inferred from the experimental results that caging-stressed ducks were in the sensitive phase in the first 5 days after caging, caging for approximately 10 days was an important transition phase, and then the duck continually adapted.