Aspergillus niger는 전분, 단백질, 셀룰로오스분해 효소 등을 생성하는 균주로 산업적으로 널리 사용되는 곰팡이이다. 본 연구에서는 누룩에서 분리한 A. niger 28균주에 대한 연구를 진행하여 국내 발표 식품 유래 균주의 유전적 다양성과 높은 셀룰로오스 분해 능력을 확인하였다. Neighbor-joining 계통 분석을 통해 모든 균주가 A. niger “aggregate” 종과 유사함을 알게 되었고 해당 결과를 바탕으로 Aspergillus의 종간 유사성을 확인하였다. 누룩유래 균주의 효소활성능을 확인하기 위해 밀기울 (...
Aspergillus niger는 전분, 단백질, 셀룰로오스분해 효소 등을 생성하는 균주로 산업적으로 널리 사용되는 곰팡이이다. 본 연구에서는 누룩에서 분리한 A. niger 28균주에 대한 연구를 진행하여 국내 발표 식품 유래 균주의 유전적 다양성과 높은 셀룰로오스 분해 능력을 확인하였다. Neighbor-joining 계통 분석을 통해 모든 균주가 A. niger “aggregate” 종과 유사함을 알게 되었고 해당 결과를 바탕으로 Aspergillus의 종간 유사성을 확인하였다. 누룩유래 균주의 효소활성능을 확인하기 위해 밀기울 (wheat bran)에서의 배양을 진행하였고 10개의 A. niger 표준 균주와 비교를 진행하였다. 그 결과, 대부분의 균주 (71.4%)가 표준 균주보다 높은 셀룰로오스 분해 능력을 보였으며 특히 KCN5와 KJC3는 높은 β-glucosidase 활성을 보였다. 두 균주의 전사체 분석을 통해 endoglucanase, cellobiohydrolase, β-glucosidase와 같은 셀룰로오스 분해 효소 유전자의 발현이 밀기울에서 높게 유지된다는 것을 확인하였다. 본 연구의 결과는 누룩유래 A. niger 균주의 유용한 효소능력, 특히 산업적으로 널리 활용될 수 있는 셀룰로오스 분해 능력을 보여주고 있다.
Aspergillus niger는 전분, 단백질, 셀룰로오스 분해 효소 등을 생성하는 균주로 산업적으로 널리 사용되는 곰팡이이다. 본 연구에서는 누룩에서 분리한 A. niger 28균주에 대한 연구를 진행하여 국내 발표 식품 유래 균주의 유전적 다양성과 높은 셀룰로오스 분해 능력을 확인하였다. Neighbor-joining 계통 분석을 통해 모든 균주가 A. niger “aggregate” 종과 유사함을 알게 되었고 해당 결과를 바탕으로 Aspergillus의 종간 유사성을 확인하였다. 누룩유래 균주의 효소활성능을 확인하기 위해 밀기울 (wheat bran)에서의 배양을 진행하였고 10개의 A. niger 표준 균주와 비교를 진행하였다. 그 결과, 대부분의 균주 (71.4%)가 표준 균주보다 높은 셀룰로오스 분해 능력을 보였으며 특히 KCN5와 KJC3는 높은 β-glucosidase 활성을 보였다. 두 균주의 전사체 분석을 통해 endoglucanase, cellobiohydrolase, β-glucosidase와 같은 셀룰로오스 분해 효소 유전자의 발현이 밀기울에서 높게 유지된다는 것을 확인하였다. 본 연구의 결과는 누룩유래 A. niger 균주의 유용한 효소능력, 특히 산업적으로 널리 활용될 수 있는 셀룰로오스 분해 능력을 보여주고 있다.
Aspergillus niger is a widely used industrial fungus due to their enzyme producing abilities, such as amylolytic, proteolytic, and cellulolytic enzymes. In this study, we obtained 28 strains of A. niger from traditional Korean fermentation starter, nuruk, which has been known as a mixed culture of e...
Aspergillus niger is a widely used industrial fungus due to their enzyme producing abilities, such as amylolytic, proteolytic, and cellulolytic enzymes. In this study, we obtained 28 strains of A. niger from traditional Korean fermentation starter, nuruk, which has been known as a mixed culture of enzymatic filamentous fungi and yeasts. By neighbor-joining phylogenetic analysis using internal transcribed spacer (ITS) region, partial β-tubulin (benA) and calmodulin gene (CaM) sequences, all of strains were identified as A. niger “aggregate” species. The amylase, protease, and cellulase production on solid wheat bran media were investigated and compared with ten reference strains of A. niger. Amylase production was measured through α- and glucoamylase assay and 17 (60.7%) strains showed high amylolytic ability. Protease production measured at various pH values (pH 3.0, 5.0, 7.0, 9.0) was highest at pH 3.0 ranged from 8.6 unit/g to 37.3 unit/g. Endoglucanase, β-glucosidase, and filter paper activity were assayed to measure cellulase production of Korean strains. Most strains (71.4%) showed higher cellulolytic activity than reference strains and particularly two strains (KJC3 and KCN5) showed high β-glucosidase activity. We performed RNA sequencing and qRT-PCR to confirm transcriptional responses of KJC3, KCN5 and reference strain N402 after exposure to wheat bran as sole carbon source. Nine hours after exposed to wheat bran, cellulolytic gene and other carbohydrate-active enzyme gene expression was highly induced. Particularly, glucanase (ATCC64974_49900 and _104930) and β-glucosidase genes (ATCC64974_13640 and _4700) of two strains were highly expressed approximately 3 to 14-fold compared to N402. And other types of CAZymes (xylanase, arabinofuranosidase and galactosidase) genes are also highly expressed as response to wheat bran as sole carbon source. Our result showed remarkable enzyme production, particularly cellulase production of A. niger strains from nuruk which could have a practical application in industry.
Aspergillus niger is a widely used industrial fungus due to their enzyme producing abilities, such as amylolytic, proteolytic, and cellulolytic enzymes. In this study, we obtained 28 strains of A. niger from traditional Korean fermentation starter, nuruk, which has been known as a mixed culture of enzymatic filamentous fungi and yeasts. By neighbor-joining phylogenetic analysis using internal transcribed spacer (ITS) region, partial β-tubulin (benA) and calmodulin gene (CaM) sequences, all of strains were identified as A. niger “aggregate” species. The amylase, protease, and cellulase production on solid wheat bran media were investigated and compared with ten reference strains of A. niger. Amylase production was measured through α- and glucoamylase assay and 17 (60.7%) strains showed high amylolytic ability. Protease production measured at various pH values (pH 3.0, 5.0, 7.0, 9.0) was highest at pH 3.0 ranged from 8.6 unit/g to 37.3 unit/g. Endoglucanase, β-glucosidase, and filter paper activity were assayed to measure cellulase production of Korean strains. Most strains (71.4%) showed higher cellulolytic activity than reference strains and particularly two strains (KJC3 and KCN5) showed high β-glucosidase activity. We performed RNA sequencing and qRT-PCR to confirm transcriptional responses of KJC3, KCN5 and reference strain N402 after exposure to wheat bran as sole carbon source. Nine hours after exposed to wheat bran, cellulolytic gene and other carbohydrate-active enzyme gene expression was highly induced. Particularly, glucanase (ATCC64974_49900 and _104930) and β-glucosidase genes (ATCC64974_13640 and _4700) of two strains were highly expressed approximately 3 to 14-fold compared to N402. And other types of CAZymes (xylanase, arabinofuranosidase and galactosidase) genes are also highly expressed as response to wheat bran as sole carbon source. Our result showed remarkable enzyme production, particularly cellulase production of A. niger strains from nuruk which could have a practical application in industry.
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