유청단백질의 분리 및 단백질 분해 효소에 의한 유청단백질의 가수분해 양상

렌친핸드(Renchinkhand),배형철(Hyoung Churl Bae),정석근(Seok Geun Jeong),남명수(Myoung Soo Nam) 충남대학교 농업과학연구소 2012 농업과학연구 Vol.39 No.4

The aim of this study was to introduce a simple method for isolation of α-lactalbumin, β-lactoglobulin and bovine serum albumin from cow’s milk, and peptides produced by enzymatic hydrolysis of α-lactalbumin, β-lactoglobulin and bovine serum albumin with alcalase. Whey protein were precipitated from whey by ammonium sulfate and, α-lactalbumin and β-lactoglobulin were isolated using Hi Prep 26/60 Sephacryl S-100 column gel filtration chromatography. Bovine serum albumin and β-lactoglobulin were isolated by Mono-Q 5/50 GL column anion exchange chromatography of the 50% Ammonium Sulfate-supernatant. Isolated whey proteins were hydrolyzed by proteolytic alcalase. Tricine SDS-PAGE and reverse-phase HPLC analyses revealed that almost hydrolyzed all the α-lactalbumin, β-lactoglobulin and bovine serum albumin with alcalase. Molecular weight of various peptides derived from alcalase hydrolysate were small molecular weight than 3.5 kDa.

강황을 첨가한 발효유의 발효특성과 면역조절 효과

렌친핸드(Renchinkhand Gereltuya),손지윤(Ji Yoon Son),어르가말(Urgamal Magsar),백승희(Seung-Hee Paik),이조윤(Jo Yoon Lee),남명수(Myoung Soo Nam) 한국생명과학회 2015 생명과학회지 Vol.25 No.1

강황 (Curcuma longa L.)은 건강기능성 식품의 훌륭한 소재로 전통적인 약용식물이다. 강황의 주요성분인 안토시아닌계인 curcumin은 항균작용, 항암작용, 항산화작용 등 다양한 효능이 있는 것으로 알려져있다. 본 연구는 혼합유산균을 스타터로 사용하여 강황 분말을 첨가한 발효유를 제조하여 발효유의 이화학적 특성 및 항염증 활성을 연구하였다. 발효유의 발효특성은 대조군에 비해 강황 첨가군에서 유산균의 성장이 현저히 빠르게 나타나 pH는 감소되었고 산도는 증가하였다. 또한 우유단백질의 분해도 부분적으로 일어났고 유기산은 lactic acid와 acetic acid가 높게 생성되었다. 기호도는 강황 특유의 쓴맛과 강한 향으로 인해 대조군에 비해 낮았다. 강황 발효유 배양액을 처리한 RAW 264.7 세포주에서 세포독성의 영향은 나타나지 않았다. 염증성 싸이토카인으로 TNF-α와 IL-6는 강황 처리군이 대조군에 비해 현저히 강하게 발현되었다. 또한 NO의 생성은 강황 처리군이 대조군에 비해 현저히 높았다. 이러한 연구결과는 강황 첨가가 유산균의 성장을 촉진시켜 발효유 제조에 도움을 주고, 염증활성을 조절하므로 강황을 이용한 발효유, 음료 제품 및 다양한 기능성 식품 소재로 활용할 수 있는 가능성을 시사하고 있다. Curcuma longa L. (CL), a traditional medicinal plant, is well known as a functional food ingredient. The major component of CL is a curcumin of anthocyanin family that has multi-functions such as antimicrobial, anticancer, and antioxidant activity. In this study, fermented milk containing CL was prepared using a mixed strain culture (Bifidobacterium bifidus, Streptococcus thermophilus, Lactobacillus acidophilus), and its physicochemical properties were characterized. In addition, inflammatory cytokine-modulating effects of the fermented milk were also investigated. As regards the properties of fermented milk, the growth rate of lactic acid bacteria in fermented milk containing CL was found to be remarkably more rapid than control. During fermentation, caseins and whey proteins were observed to be partially hydrolyzed, and lactic acid and acetic acid were produced in larger amounts than in the control. The sensory score of fermented milk containing CL was lower than control, owing to its bitter taste and strong flavor. RAW 264.7 cells treated with CL fermented milk supernatant showed no cytotoxicity. Inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) were significantly produced by fermented milk with CL, compared to control. The secretion of nitric oxide (NO) from RAW 264.7 cells significantly increased relative to the control. Results from the present study suggested that CL could be used as a natural immunomodulating ingredient for making yogurts, beverages, and other products.

Biotransformation of Major Ginsenoside Rb₁ to Rd by Dekkera anomala YAE-1 from Mongolian Fermented Milk (Airag)

( Gereltuya Renchinkhand ),( Soo-Hyun Cho ),( Young W. Park ),( Gyu-yong Song ),( Myoung Soo Nam ) 한국미생물 · 생명공학회 2020 Journal of microbiology and biotechnology Vol.30 No.10

Dekkera anomala YAE-1 strain separated from “airag” (Mongolian fermented mare’s milk) produces β-glucosidase, which can convert ginsenoside Rb₁ from Panax ginseng. Ginseng- derived bioactive components such as ginsenoside Rb₁ have various immunological and anticancer activities. Airag was collected from five different mare milk farms located near Ulaanbaatar, Mongolia. YAE-1 strains were isolated from airag to examine the hydrolytic activities of β-glucosidase on Korean Panax ginseng using an API ZYM kit. Supernatants of selected cultures having β-glucosidase activity were examined for hydrolysis of the major ginsenoside Rb₁ at 40°C, pH 5.0. The YAE-1 strain was found to be nearly identical at 99.9% homology with Dekkera anomala DB-7B, and was thus named Dekkera anomala YAE-1. This strain exerted higher β-glucosidase activity than other enzymes. Reaction mixtures from Dekkera anomala YAE-1 showed great capacity for converting ginsenoside Rb₁ to ginsenoside Rd. The β-glucosidase produced by Dekkera anomala YAE-1 was able to hydrolyze ginsenoside Rb₁ and convert it to Rd during fermentation of the ginseng. The amount of ginsenoside Rd was highly increased from 0 to 1.404 mg/ml in fermented 20% ginseng root at 7 days.

Characterization of Paenibacillus sp. MBT213 Isolated from Raw Milk and Its Ability to Convert Ginsenoside Rb1 into Ginsenoside Rd from Panax ginseng

Gereltuya Renchinkhand,Soo Hyun Cho,Magsar Urgamal,Young W Park,Joong Hyeon Nam,Hyung Churl Bae,Gyu Yong Song,Myoung Soo,Nam 한국축산식품학회 2017 한국축산식품학회지 Vol.37 No.5

This study was conducted to isolate and characterize Paenibacillus sp. MBT213 possessing β-glucosidase activity from raw milk, and examine the enzymatic capacity on the hydrolysis of a major ginsenoside (Rb1). Strain MBT213 was found to have a high hydrolytic ability on ginsenoside Rb1 by Esculin Iron Agar test. 16S rDNA analysis revealed that MBT213 was Paenibacillu sp. Crude enzyme of MBT213 strain exhibited high conversion capacity on ginsenoside Rb1 into ginsenoside Rd proven by TLC and HPLC analyses. The API ZYM kit confirmed that Paenibacillu sp. MBT213 exerted higher β-glucosidase and β-galactosidase activity than other strains. Optimum pH and temperature for crude enzyme were found at 7.0 and 35ºC in hydrolysis of ginsenoside Rb1. After 10 d of optimal reaction conditions for the crude enzyme, ginsenoside Rb1 fully converted to ginsenoside Rd. Ginseng roots (20%) were fermented for 14 d, and analyzed by HPLC showed that amount of ginsenoside Rb1 significantly decreased, while that of ginsenoside Rd was significantly increased. The study confirmed that the β-glucosidase produced by Paenibacillus sp. MBT213 can hydrolyze the major ginsenoside Rb1 and convert to Rd during fermentation of the ginseng. The β-glucosidase activity of this novel Paenibacillus sp. MBT213 strain may be utilized in development of variety of health foods, dairy foods and pharmaceutical products.

Alcalase에 의한 유청단백질 가수분해물의 항원성 저감 효과

유재민(Renchinkhand),렌친핸드(Hyoung Churl Bae),정석근(Seok Geun Jeong),백승희(Seung-Hee Paik),남명수(Myoung Soo Nam) 충남대학교 농업과학연구소 2013 농업과학연구 Vol.40 No.4

The aim of this study was to produce enzymatic hydrolysis of α-LA, β-LG and BSA with alcalase for the possible application of hypoallergenic foods toward cow’s milk allergenic infant. The molecular weights of most of the peptides in hydrolysates from α-LA, β-LG and BSA by alcalase were below 3,000 dalton. Antigenesity of α-LA, β-LG and BSA hydrolysates to rabbit anti-α-LA antiserum, β-LG antiserum and BSA antiserum were remarkably decreased by more than 10-3 at 20% inhibitionrate. Antigenesity of polyvalent antigenic peptide in α-LA, β-LG and BSA hydrolysates to specific rabbit anti-α-LA antiserum, β-LG antiserum and BSA antiserum was determined by PCS test using guina-pig. Hydrolysates of α-LA, β-LG and BSA with less than 3,000 dalton did not show polyvalent antigenic reaction against rabbit antiserum. Hydrolysates of α-LA, β-LG and BSA could be a source for the manufacturing of hypoallergenic food.

초유에 함유된 성장인자와 기능: 총설

렌친핸드 ( Gereltuya Renchinkhand ),손지윤 ( Ji Yoon Son ),남명수 ( Myoung Soo Nam ) 한국유가공기술과학회 2016 Journal of Dairy Science and Biotechnology (JMSB) Vol.34 No.1

젖소 초유에는 성장인자가 풍부하게 함유되어 있는데, 상처 치유에 중요한 역할을 하고, 초유의 생리활성 기능을 담당하고 있다. Tyrosine kinase receptor의 활성을 유도하는 성장인자가 특이적으로 관여하여 세포의 분화, 면역기능, 신경기능 등 세포간 상호작용에 관여하는 EGFR(상피증식인자 수용체)와 FGFR(섬유아세포 증식인자)가 있다. 또한 VEGFR(혈관내피 증식인자)와 PDGF(혈소판유래 증식인자)도 존재한다. 조직회복을 위한 각질세포 분화와 세포의 이행에 성장인자가 상승효과를 나타내었고, 초유 또는 초유에 포함된 성장인자 peptide들은 장관질환 치료에 효과가 있으므로 치료제로 이용 가능성을 보여주었다. Colostrum, a nutrient-rich fluid produced by female mammals after giving birth, is the specific initial diet of mammalian neonates. Colostrum is important for the nutrition, growth, and development of newborn infants and contributes to the immunologic defense of neonates. It contains immunoglobulins, antimicrobial peptides, such as lactoferrin and lactoperoxidase, and other bioactive molecules, including growth factors, such as IGF (insulin-like growth factor), EGF (epithermal growth factor), TGF-β (transforming growth factor), and FGF (fibroblast growth factor). Bovine colostrum is a rich source of growth factors, which play a central role in wound healing. The biological activities of colostrum emphasize the relevance of the synergistic activity of growth factors to stimulate keratinocyte proliferation and migration, which are essential for tissue repair. Colostrum increases the expression of early differentiation markers, such as keratin 1 and 10 and involucrin, and late differentiation markers, including loricrin and filaggrin. Additionally, colostrum increases granulation tissue volume in the dermis, suggesting that it has a beneficial effect on wound healing. The therapeutic use of colostrum or individual peptides present in colostrum has a positive and curative influence on various gastrointestinal diseases.

Biological properties of fermented milk with fortified whey protein

김기환,라석한,Renchinkhand Gereltuya,기우진,남명수,김완섭 충남대학교 농업과학연구소 2023 Korean Journal of Agricultural Science Vol.50 No.2

As a byproduct obtained from cheese manufacture, whey protein was developed as a functional food that contains multi-functional proteins. In this study, the biochemical activity of fermented milk prepared by fortifying whey protein with excellent physiological activity was investigated. Immunoglobulin (IgG) content was higher in 10% fortified whey protein fermented milk than in the control. The viable cell counts were 20% higher in the fermented milk with 10% fortified whey protein than in the control group. The antibacterial effect of 10% fortified whey protein fermented milk compared to the control group was shown to be effective against four pathogenic microorganisms, Escherichia coli (KCTC1039), Pseudomonas aeruginosa 530, Salmonela Typhimurium (KCTC3216), and Staphylococcus aureus (KCTC1621). The antioxidant effect by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities wasincreased two-fold in 10% fortified whey protein fermented milk compared to the control. The 10% fortified whey protein fermented milk inhibited the expression of the inflammatory cytokines (interleukin [IL]-6, tumor necrosis factor [TNF]-α, and induced nitric oxide synthase [iNOS]) in a concentration-dependent manner. In a piglets feeding test, the weight gain with 10% fortified whey protein fermented milk was increased by 18% compared to the control group, and no diarrhea symptoms appeared. Our results clearly demonstrated that 10% fortified whey protein fermented milk could be a useful functional ingredient for improving health.

Cytokine modulation in Raw 264.7 macrophages treated with ginseng fermented by Penibacillus MBT213

손지윤,Gereltuya Renchinkhand,배형철,백승희,이조윤,남명수 충남대학교 농업과학연구소 2018 Korean Journal of Agricultural Science Vol.45 No.4

The fermentation of Panax ginseng yields many compounds including ginsenosides that have various biological functions. The objective of this study was to investigate the modulation of nitric oxide (NO), Interleukin (IL)-6 and tumor necrosis factor (TNF)-α in Raw 264.7 cells treated with ginseng fermented by Penibacillus MBT213. Nitric oxide production in the Raw 264.7 cells treated for 24 hours with fermented ginseng at 3, 7, and 14 days after the treatment decreased to 74, 43, and 36%, respectively, compared with the positive control. The production of IL-6 was inhibited in all the cells treated with fermented ginseng at 3, 7, and 14 days after the treatment except for the positive control. The TNF-α production in the Raw 264.7 cells treated with fermented ginseng for 6 hours at 3, 7, and 14 days after the treatment was about 40,000, 85,000 and 65,000 pg/mL, respectively. Moreover, the TNF-α production in the Raw 264.7 cells treated with fermented ginseng for 24 hours at 7 and 14 days after the treatment was about 160,000 and 180,000 pg/mL, respectively. However, TNF-α production was inhibited in the Raw 264.7 cells at 6 and 12 hours after the treatment with fermented ginseng. herefore, it was confirmed that the immunological activity of the Raw 264.7 macrophages was affected by the treatment with fermented ginseng. It was concluded that ginseng fermented by Paenibacillus MBT213 possesses a potential anti-inflammatory activity and could be used as an ingredient in functional foods and pharmaceutical products.

Antioxidant, antibacterial, and antiinflammatory effects of yoghurt made with vitamin tree (Hippophae rhamnoides L.) fruit powder

Byung Bae Park,Gereltuya Renchinkhand,Woo Jin Ki,Jong Woo Choi,Myoung Soo Nam 충남대학교 농업과학연구소 2022 Korean Journal of Agricultural Science Vol.49 No.4

Effect of lactoferrin hydrolysates on inflammatory cytokine modulation in HEK- 293, RBL-2H3, and HMC-1 cells

손지윤,배형철,GERELTUYA RENCHINKHAND,남명수,김완섭 충남대학교 농업과학연구소 2020 Korean Journal of Agricultural Science Vol.47 No.1

Lactoferrin (LF) is an iron-binding glycoprotein that is present in colostrum, milk, and other body secretions. The objective of this study was to investigate the effects of lactoferrin hydrolysates (LHs) on the production of immunomodulatory factors, including inflammatory related cytokines. The nuclear factor (NF)-κB reporter assay using human embryonic kidney 293 cells (HEK-293) revealed that NF-κB activity was significantly decreased by 1, 50, and 100 μg/mL of LH and the fractions above and below the 10 kDa LH. The mRNA expression of interferon (IFN)-γ in rat basophilic leukemia mast cells (RBL-2H3) treated with the fraction above the 10 kDa LH decreased in a dose-dependent manner, but the cells treated with LH and the fraction below the 10 kDa LH showed an increased expression of IFN-γ in a dose-dependent manner. The level of cyclooxygenase (COX)-2 expression decreased dosedependently in RBL-2H3 cells treated with LH and the fraction above the 10 kDa LH, but the cells treated with the fraction below the 10 kDa LH showed an increased COX-2 expression in a dose-dependent manner. The mRNA expression of interleukin (IL)-4) was dose-dependently decreased by the fraction below the 10 kDa LH in human mast cells (HMC-1). The mRNA expressions of tumor necrosis factor (TNF)-α and IL-6 were significantly dose-dependently decreased by the fractions above and below the 10 kDa LH, but was dose-dependently increased by LH. The production of IL-4 was a little increased by the fraction above the 10 kDa LH compared to the positive control, but was decreased with LH and the fraction below the 10 kDa LH in HMC-1 cells. It was concluded that LF hydrolysates had an immunomodulating effect on anti-, pro-inflammatory and anti-allergic reactions.