제주산 보리 및 추출된 β-Glucan의 이화학적 특성

김효진,김현정 한국산업식품공학회 2015 산업 식품공학 Vol.19 No.2

본 연구에서는 제주 지역에서 재배하고 생산되는 보리의 이화학적 특성을 분석하고, 수용성 식이섬유인 β-glucan을 추출하여 타 지역에서 재배된 보리와 비교, 분석하여 제주 보리가 기능성을 갖춘 제품으로 활용될 수 있는 자료를 제 시하고자 하였다. 제주 지역에서 재배된 보리는 전반적으 로 전분 함량과 식이섬유 함량이 높았으며, 특히 수용성 식이섬유인 β-glucan의 함량이 높은 것으로 나타났다. 제주 맥주보리는 타 지역 보리와 유사한 성분 함량을 갖는 것으 로 나타났으며 도정유무가 전분, 식이섬유와 β-glucan 함량 에 영향을 주는 것으로 나타났다. 보리의 점도는 가열 처 리에 의해 상당량 증가하였는데 상온에서는 제주쌀보리가 102.5 cP의 높은 점도를 나타내었고 가열 후에는 4563.3 cP로 제주청보리가 가장 높은 점도를 나타내어 타 지역 보 리에 비해 점도가 높게 나타났다. 또한 도정 유무가 점도 에 영향을 미쳤는데, 도정되지 않은 제주맥주보리의 점도 가 도정된 제주맥주보리에 비해 현저히 감소하였다. 열수 와 효소를 이용하여 보리로부터 비교적 높은 수율의 β- glucan을 추출하였다(70-80%). 추출된 β-glucan의 재용해율 은 보리의 품종에 따라 다르게 나타났는데, 검정보리와 제 주쌀보리의 재용해율이 높은 것으로 나타났다. 도정되지 않은 제주맥주보리는 가장 낮은 재용해율을 보여 도정이 추출된 β-glucan의 물리적 특성에 영향을 미치는 것으로 나타났다. β-Glucan을 구성하는 glucose들간의 β(1→3):β(1 →4) 결합비는 1:2.3-4.49의 범위에서 측정되었으며, 정제 정도만이 β(1→3):β(1→4) 결합비에 영향을 미치는 것이 아 니라 도정율과 같은 다른 요인들도 영향을 미칠 수 있다는 것을 확인하였고, 구체적으로 어떠한 요인들이 β(1→3):β(1 →4) 결합비에 영향을 미칠 수 있는지에 대한 연구가 더 필요한 것으로 보인다. 본 연구를 통하여 제주 지역에서 재배된 보리의 일반성분과 온도에 따른 점도 특성, 추출된 β-glucan의 이화학적 특성의 결과를 바탕으로 제주 보리를 활용한 새로운 기능소재 및 제품 개발에 이용 가능성이 높 을 것으로 기대되었다. This study investigated the physicochemical properties of barley varieties grown in Jeju along with the characteristics of soluble dietary fiber β-glucan extracted from barley. Jeju naked barley and blue barley contained greater amounts of starch and total dietary fiber than other varieties of barley, and their β-glucan contents were 8.63 and 7.02%, respectively. Unhulled barley, black barley, and Jeju beer barley contained 4.49-5.86% of β-glucan. The viscosity of barley solution greatly increased by heating and the viscosity values of Jeju naked barley and blue barley at heating were 2350.0 and 4563.3 cP, which were the highest. When compared to unpolished and polished Jeju beer barley, the viscosity increased by polishing. β-Glucan was efficiently extracted from different barley varieties where extraction yield ranged from 70.12 to 85.23%. The resolubilities of extracted β-glucans were significantly different depending on the varieties of barley (p<0.05). The ratio of β(1→3) to β(1→4) linkage composed of β-glucan was measured in the range of 1:2.3-4.49. These results suggested that the barley grown in Jeju containing a high amount of β-glucan presented different physicochemical properties and could possibly be applied to develop various types of food products with beneficial health functions.

Expression and characterization of a novel microbial GH9 glucanase, IDSGLUC9-4, isolated from sheep rumen

Zhu Yongzhen,Bai Shuning,Li Nuo,Wang Jun-Hong,Wang Jia-Kun,Wang Qian,Wang Kaiying,Zhang Tietao 아세아·태평양축산학회 2024 Animal Bioscience Vol.37 No.9

Objective: This study aimed to identify and characterize a novel endo-β-glucanase, IDSGLUC9-4, from the rumen metatranscriptome of Hu sheep.Methods: A novel endo-β-glucanase, IDSGLUC9-4, was heterologously expressed in <i>Escherichia coli</i> and biochemically characterized. The optimal temperature and pH of recombinant IDSGLUC9-4 were determined. Subsequently, substrate specificity of the enzyme was assessed using mixed-linked glucans including barley β-glucan and Icelandic moss lichenan. Thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), matrix assisted laser desorption ionization time of flight mass spectrometry analyses were conducted to determine the products released from polysaccharides and cello-oligosaccharides substrates.Results: The recombinant IDSGLUC9-4 exhibited temperature and pH optima of 40°C and pH 6.0, respectively. It exclusively hydrolyzed mixed-linked glucans, with significant activity observed for barley β-glucan (109.59±3.61 μmol/mg min) and Icelandic moss lichenan (35.35±1.55 μmol/mg min). TLC and HPLC analyses revealed that IDSGLUC9-4 primarily released cellobiose, cellotriose, and cellotetraose from polysaccharide substrates. Furthermore, after 48 h of reaction, IDSGLUC9-4 removed most of the glucose, indicating transglycosylation activity alongside its endo-glucanase activity.Conclusion: The recombinant IDSGLUC9-4 was a relatively acid-resistant, mesophilic endo-glucanase (EC 3.2.1.4) that hydrolyzed glucan-like substrates, generating predominantly G3 and G4 oligosaccharides, and which appeared to have glycosylation activity. These findings provided insights into the substrate specificity and product profiles of rumen-derived GH9 glucanases and contributed to the expanding knowledge of cellulolytic enzymes and novel herbivore rumen enzymes in general. Objective: This study aimed to identify and characterize a novel endo-β-glucanase, IDSGLUC9-4, from the rumen metatranscriptome of Hu sheep. Methods: A novel endo-β-glucanase, IDSGLUC9-4, was heterologously expressed in Escherichia coli and biochemically characterized. The optimal temperature and pH of recombinant IDSGLUC9-4 were determined. Subsequently, substrate specificity of the enzyme was assessed using mixed-linked glucans including barley β-glucan and Icelandic moss lichenan. Thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), matrix assisted laser desorption ionization time of flight mass spectrometry analyses were conducted to determine the products released from polysaccharides and cello-oligosaccharides substrates. Results: The recombinant IDSGLUC9-4 exhibited temperature and pH optima of 40°C and pH 6.0, respectively. It exclusively hydrolyzed mixed-linked glucans, with significant activity observed for barley β-glucan (109.59±3.61 μmol/mg min) and Icelandic moss lichenan (35.35±1.55 μmol/mg min). TLC and HPLC analyses revealed that IDSGLUC9-4 primarily released cellobiose, cellotriose, and cellotetraose from polysaccharide substrates. Furthermore, after 48 h of reaction, IDSGLUC9-4 removed most of the glucose, indicating transglycosylation activity alongside its endo-glucanase activity. Conclusion: The recombinant IDSGLUC9-4 was a relatively acid-resistant, mesophilic endo-glucanase (EC 3.2.1.4) that hydrolyzed glucan-like substrates, generating predominantly G3 and G4 oligosaccharides, and which appeared to have glycosylation activity. These findings provided insights into the substrate specificity and product profiles of rumen-derived GH9 glucanases and contributed to the expanding knowledge of cellulolytic enzymes and novel herbivore rumen enzymes in general.

β-(1,3)-Glucan Isolated from Agrobacterium Species Induces Maturation of Bone Marrow-derived Dendritic Cells and Drives Th1 Immune Responses

변의백,박상현,성낙윤,변의홍,박철환 한국식품과학회 2015 Food Science and Biotechnology Vol.24 No.4

The β-(1,3)-glucan originally isolated from Agrobacterium species was investigated for prompting maturation of dendritic cells (DCs) and driving Th1 immune responses. Bone-marrow derived DCs separated from mice were analyzed for augmentation of cell surface molecule (CD80, CD86, and major histo-compatability complex (MHC) class I/II) expression and pro-inflammatory cytokine production (tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6). β-Glucan functionally induced DCs activation via augmentation of CD80, CD86, and MHC class I/II expression and cytokine production (TNF-α, IL-1β, and IL-6). β-Glucan induced secretion of IL-12p70, but not IL-10. Both mitogen-activated protein kinases and nuclear factor (NF)-κB signaling mediated production of pro-inflammatory cytokine induced by β- glucan. β-Glucan-treated DCs exhibited accelerated proliferation of murine splenocytes with increased levels of interferon (IFN)-γ. β-Glucan regulates innate and adaptive immunity via DCs activation and Th1 polarization of immune responses.

반응표면분석법을 이용한 흰목이버섯의 기능성 성분 추출 조건 최적화

홍민,최다혜,한준희,권태형,이선엽,이용진,유근형 한국버섯학회 2020 한국버섯학회지 Vol.18 No.4

Snow fungus (Tremella fuciformis) with functional contents has satisfactory effects on various diseases, including atherosclerosis, high cholesterol, healthy skin, cancer, diabetes, and anti-inflammation. In this study, the extraction yield and functional contents (ergothioneine and β-glucan) of white jelly fungus were compared based on the extraction conditions using response surface methodology. Results revealed the extraction conditions for optimization of the dependent variables to be 60oC and 4.33 h, when 16.6 mg/mL of sample concentration was used. Under these conditions, the extraction yield was 24.9%, including ergothioneine (66.8 μg/g) and β-glucan (34.9 g/100 g). These results can be useful in understanding the functional ingredients and mass extraction process in mushroom.

Isolation and Characterization of Saccharomyces cerevisiae Mutants Deficient in (1→3)-β-D-Glucan Synthase

Song, M. R.,Lee, D. W.,Park, S. W.,Bae, K. S.,Park, H. M. 한국산업미생물학회 1992 한국미생물·생명공학회지 Vol.20 No.6

본인 등은 베타-1,3-글루칸 합성효소능이 낮은 Saccharomyces cerevisiae의 조건성 돌연변이주들을 분리하여 그 특성을 조사하였다. 이 돌연변이주들은 비 허용온도인 37℃에서 삼투감수성을 나타나며, 세포벽 성분 중 알칼리 비용해성 글루칸의 함량이 낮았다. 베타-1,3-글루칸 합성효소능의 결함원인을 조사한 결과, 효모의 베타-1,3-글루칸 합성효소를 구성하는 촉매성분(catalytic component)과 GTP-결합성분 두 가지 중 촉매성분의 결함이 이 돌연변이주들의 베타-1,3-글루칸 합성효소능의 손상원인인 것으로 추정되었다. We have isolated conditional lethal mutants of Saccharomyces cerevisiae which are low in (1→3)-β-D-glucan synthase activity. These mutants were osmotic sensitive at nonpermissive temperature (37℃) and showed a decreased level of alkali-insoluble cell wall glucan. The decrease in (1→3)-β-D-glucan synthase activity of the mutants appeared to be mainly due to the defect in catalytic component rather than in GTP-binding component.

Isolation and Characterization of Saccharomyces cerevisiae Mutants Deficient in (1→3)-β-D-Glucan Synthase

Song, M. R.,Lee, D. W.,Park, S. W.,Bae, K. S.,Park, H. M. 충남대학교 생물공학연구소 1993 생물공학연구지 Vol.3 No.-

본인 등은 베타-1,3-글루칸 합성효소능이 낮은 Saccharomyces cerevisiae의 조건성 돌연변이주들을 분리하여 그 특성을 조사하였다. 이 돌연변이주들은 비허용온도인 37℃ 에서 삼투감수성을 나타나며, 세포벽성분 중 알칼리 비용해성 글루칸의 함량이 낮았다. 베타-1,3-글루칸 합성효소능의 결함원인을 조사한 결과, 효모의 베타-1,3-글루칸 합성효소를 구성하는 촉매성분(catalytic component)과 GTP-결합성분 두 가지 중 촉매성분의 결함이 이 돌연변이주들의 베타-1,3-글루칸 합성효소능의 손상원인인 것으로 추정되었다. We have isolated conditional lethal mutants of Saccharomyces cerevisiae which are low in (1→3)-β-D-glucan synthase activity. These mutants were osmotic sensitive at nonpermissive temperature (37℃)and showed a decreased level of alkali-insoluble cell wall glucan. The decrease in (1→3)-β-D-glucan synthase activity of the mutants appeared to be mainly due to the defect in catalytic component rather than in GTP-binding component.

Enzyme activity and expression pattern of intra- and extracellular chitinase and β-1,3-glucanase of Wickerhamomyces anomalus EG2 using glycol chitin and glucan-containing high polymer complex

Hong, S.H.,Song, Y.S.,Seo, D.J.,Kim, K.Y.,Jung, W.J. Elsevier 2017 INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES Vol.105 No.1

We investigated cell growth and activity of intra- and extracellular chitinase, β-1,3-glucanase, and chitin deacetylase with SDS@?PAGE by incubating W. anomalus EG2 in PDB and YPD media for 24h in presence of different concentrations (0%, 0.1%, 0.3%, and 0.5%) of colloidal chitin. Maximum cell growth was observed in both PDB and YPD media without colloidal chitin. In the absence of colloidal chitin, maximum extracellular β-1,3-glucanase activity of 32.96 and 47.28 units/mL was reported at 18h in PDB medium and 6h in YPD medium, respectively. In addition, extracellular chitinase was unaffected by various concentrations of carboxymethyl chitin in both PDB and YPD media. In the absence of colloidal chitin, maximum intracellular chitinase activity was indicated to be 9.82 and 9.86 units/mg protein in PDB and YPD media, respectively. Maximum intracellular β-1,3-glucanase activity reported was 17.34 units/mg protein in PDB medium containing 0.5% colloidal chitin and 15.0 units/mg protein in YPD medium containing 0.3% colloidal chitin. Five major isozymes, GN1, GN2, GN3, GN4, and GN5, of intracellular β-1,3-glucanase were detected with glucan-containing high polymer complex as a substrate with or without colloidal chitin.

Characterization of Exopolysaccharide (EPS) Produced by Weissella hellenica SKkimchi3 Isolated from Kimchi

Min Ju Kim,Ha Na Seo,황태식,Sung Hun Lee,박두현 한국미생물학회 2008 The journal of microbiology Vol.46 No.5

Weissella hellenica SKkimchi3 produces the higher exopolysaccharide (EPS) on sucrose than lactose, glucose, and fructose at pH 5 and 20°C. Sucrose was exclusively used to cultivate SKkimchi3 in all experiments base on the EPS production tests. The molecular mass of EPS, as determined by gel permeation chromatography, was 203,000. ¹H and ¹³C NMR analysis indicated that the identity of EPS may be a glucan. When EPS, starch, and cellulose was treated with α-amylase, glucoamylase, glucosidase, and cellulase, glucose was produced from starch and cellulose but was not produced from EPS. Based on HPLC analysis, elemental analysis, ¹H and ¹³C NMR analysis, and enzymatic hydrolysis tests, EPS was estimated to be a glucan. EPS suspension was not precipitated even by centrifugation at 10,000×g for 60 min, and EPS made the fermented milk and bacterial culture viscous.

β(1-3)-D-glucan affects adipogenesis, wound healing and inflammation

Vaclav Vetvicka,Jana Vetvickova 경희대학교 융합한의과학연구소 2011 Oriental Pharmacy and Experimental Medicine Vol.11 No.3

Numerous types of β(1-3)-D-glucans have been isolated from almost every species of yeast, grain, and fungi. These products have been extensively studied for their immunological and pharmacological effects. In this paper we evaluated the possibility whether individual β(1-3)-D-glucans will have an activity in less studied areas such as adipogenesis and inflammation. Our results showed that of the tested β(1-3)-D-glucans, yeast-derived insoluble Glucan #300, strongly inhibited adipogenic differentiation,supported wound healing and significantly lowered skin irritation. The remaining β(1-3)-D-glucans were significantly less active. Taken together, our study showed that with respect to natural β(1-3)-D-glucans, there is a clear yes-or-no effect suggesting that highly purified and highly active β(1-3)-D-glucans will have pleiotropic biological impact, whereas poorly isolated and/or less active β(1-3)-D-glucans will have only mediocre properties.

유산균 발효한 현미상황버섯균사체 추출물의 β-Glucan 함량 및 주름개선 효과

김재원,안정모,권오준,김선희 한국피부과학연구원 2016 아시안뷰티화장품학술지 Vol.14 No.2

Purpose: This study aimed to investigate the potential of brown rice Phellinus linteus as an anti-wrinkle material for cosmetics through examining anti-wrinkle effects. Methods: To check the anti-wrinkle effects of brown rice Phellinus linteus mycelium, we performed various experiments such as β-glucan content, β-glucan molecular weight, cell viability, matrix metalloproteinase-1 (MMP-1) inhibitory activity, and pro-collagen synthesis assay.Results: The β-glucan contents from brown rice Phellinus linteus mycelium extracts unfermented and fermented were 8.1, 12.8 g/100 g, rose up respectively and the fermented increased 58% compared with unfermented extract. The low molecular weights of the β-glucan such as less than 2500 m/z from the fermented was measured by MALDI-TOF mass spectrometry. We found that high molecular weight of β-glucan decomposed low molecular weight through fermentation. Measuring cytotoxicity of brown rice Phellinus linteus extracts in HS27 cells, cytotoxicity of both unfermented and fermented was not observed under 1,500 μg/mL concentration. After irradiating UVB treatment, HS27 cell viability of unfermented and fermented both showed the highest effect at 800 μg/mL concentration. The inhibitory activity of collagenase (MMP-1) of unfermented and fermented both showed the highest effect at a concentration of 400 μg/mL and the inhibitory activity of collagenase (MMP-1) of the fermented at a concentration of 1000 μg/mL had similar effects with adenosine at a concentration of 400 μg/mL. The amounts of biosynthesis of pro-collagen type I of the unfermented and fermented both increased more than three times compared with those of control. Conclusion: Through results, we identified anti-wrinkle effects of the brown rice Phellinus linteus and we anticipated the brown rice Phellinus linteus develop the functional cosmetic ingredient as anti-wrinkle effects 목적: 본 연구는 주름개선 효과검정을 통해 현미상황버섯균사체의 주름개선 기능성 화장료 소재로서의 가능성을 확인하는데 그목 적이 있다. 방법: Lactobacillus plantarum 으로 현미상황버섯균사체를 배양하여 얻어진 추출물의 β-glucan 함량, β-glucan 분자량 측정, 세포독성평가, MMP-1 저해활성 측정, 콜라겐생합성 효과를 시험하였다. 결과: 현미상황버섯균사체의 β-glucan 함량 분석결과 미발효군, 발효군에서 각각 8.1, 12.8 g/100 g의 결과로 함량이 58% 증가하는 것을 입증하였으며, β-glucan의 분자량 측정을 실시한 결과, 2,500 m/z 이하의 저분자 β-glucan을 확인하여 발효공정이 고분자의 β-glucan을 저분자로 분해하는 것을 확인하였다. 추출물의 세포독성은 미발효군, 발효군 모두 2,000μ g/mL이하 농도에서 세포독성이 나타나지 않아 독성에 대한 안전성을 확인하였다. 자외선(UVB) 조사에 의한 세포생존률은 미발효군, 발효군 모두800 μg/mL 농도에서 가장 높은 세포생존율 효과를 나타냈다. Matrix metalloproteinase-1 (MMP-1)의 생성억제를 확인한 결과미발효군, 발효군 모두 400 μg/mL 이상의 농도에서 MMP-1의 생성억제의 효과가 나타내는 것을 확인하였으며, 발효군 1,000 μg/mL 이상의 농도에서는 adenosine 400 μg/mL 과 유사한 효과로 높은 저해활성을 나타내어 colalgenase 저해활성이 우수함을확인하였다. Pro-collagen type Ⅰ의 생합성량을 측정한 결과 대조군 대비 미발효군, 발효군 모두 3배 이상의 collagen 생합성 증가를 나타내었다. 결론: 위의 내용을 종합하여 주름개선용 기능성 화장료로서 현미상황버섯균사체 유산균 발효 배양물의 활용가능성을 제시하였다.