Seo, Dong-Ho
(Research Group of Healthcare, Korea Food Research Institute)
,
Cho, Eui-Sang
(Department of Bioengineering and Nano-Bioengineering, Graduate School of Incheon National University)
,
Hwang, Chi Young
(Division of Bioengineering, Incheon National University)
,
Yoon, Deok Jun
(Division of Bioengineering, Incheon National University)
,
Chun, Jeonghye
(Division of Bioengineering, Incheon National University)
,
Jang, Yujin
(Division of Bioengineering, Incheon National University)
,
Nam, Young-Do
(Research Group of Healthcare, Korea Food Research Institute)
,
Park, So-Lim
(Research Group of Healthcare, Korea Food Research Institute)
,
Lim, Seong-Il
(Research Group of Healthcare, Korea Food Research Institute)
,
Kim, Jae-Hwan
(Advanced Geomaterials R&D Department, Pohang Branch, Korea Institute of Geoscience and Mineral Resources)
,
Seo, Myung-Ji
(Department of Bioengineering and Nano-Bioengineering, Graduate School of Incheon National University)
We have isolated and identified 72 bacterial strains from four bentonite samples collected at the mining areas located in Gyeongsangbuk-do, Republic of Korea, and measured their hydrolytic enzyme (${\alpha}$-amylase, protease, and cellulase) activities to identify the isolates with indust...
We have isolated and identified 72 bacterial strains from four bentonite samples collected at the mining areas located in Gyeongsangbuk-do, Republic of Korea, and measured their hydrolytic enzyme (${\alpha}$-amylase, protease, and cellulase) activities to identify the isolates with industrial-use potential. Most of the isolates belonged to the Bacillaceae, with minor portions being from the Paenibacillaceae, Micrococcaceae, and Bacillales Family XII at the family level. Of the strains isolated, 33 had extracellular ${\alpha}$-amylase activity, 30 strains produced cellulase, and 35 strains produced protease. Strain MBLB1268, having the highest ${\alpha}$-amylase activity, was identified as Bacillus siamensis ($0.38{\pm}0.06U/ml$). Bacillus tequilensis MBLB1223, isolated from Byi33-b, showed the highest cellulase activity ($0.26{\pm} 0.04U/ml$), whereas Bacillus wiedmannii MBLB1197, isolated from Zdb130-b, exhibited the highest protease activity ($54.99{\pm}0.78U/ml$). These findings show that diverse bacteria of the Bacillaceae family adhere to and exist in bentonite and are potential sources of industrially useful hydrolytic enzymes.
We have isolated and identified 72 bacterial strains from four bentonite samples collected at the mining areas located in Gyeongsangbuk-do, Republic of Korea, and measured their hydrolytic enzyme (${\alpha}$-amylase, protease, and cellulase) activities to identify the isolates with industrial-use potential. Most of the isolates belonged to the Bacillaceae, with minor portions being from the Paenibacillaceae, Micrococcaceae, and Bacillales Family XII at the family level. Of the strains isolated, 33 had extracellular ${\alpha}$-amylase activity, 30 strains produced cellulase, and 35 strains produced protease. Strain MBLB1268, having the highest ${\alpha}$-amylase activity, was identified as Bacillus siamensis ($0.38{\pm}0.06U/ml$). Bacillus tequilensis MBLB1223, isolated from Byi33-b, showed the highest cellulase activity ($0.26{\pm} 0.04U/ml$), whereas Bacillus wiedmannii MBLB1197, isolated from Zdb130-b, exhibited the highest protease activity ($54.99{\pm}0.78U/ml$). These findings show that diverse bacteria of the Bacillaceae family adhere to and exist in bentonite and are potential sources of industrially useful hydrolytic enzymes.
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문제 정의
In this study, we isolated microorganisms in domestic bentonites to investigate cultivable microbial diversity. Furthermore, their hydrolytic enzyme activities including α-amylase, cellulase, and protease, were evaluated to provide basically scientific information and explore industrial applicability of bentonite-derived microorganisms.
대상 데이터
Four bentonite samples were collected at the Gampo-40, Youngil-33, Daebo-130, and Guryong-130 mining areas (denoted Bgp40-b, Byi33-b, Zdb130-b, and Bdb130-1, respectively) located in Gyeongsangbuk-do, Republic of Korea (Table 1). Each of bentonite samples (1 g) was crushed and suspended in distilled water (10 ml) to determine pH and NaCl concentration (%, w/v) by pH meter (Fisher Science Education, USA) and HI 96821 Refractometer (HANNA instruments, USA), respectively.
이론/모형
The evolutionary distances were determined using the Kimura two-parameter model [13]. The Phylogenetic tree of the 16S rRNA gene sequences of the isolated strains and related taxa was constructed using MEGA7 program [14] based on the neighbor-joining (NJ) algorithm [15].
The activities of α-amylase and cellulase in the primary selected isolates was measured by the DNS (3,5-dinitrosalicylicacid) method.
net/eztaxon/) [12]. The evolutionary distances were determined using the Kimura two-parameter model [13]. The Phylogenetic tree of the 16S rRNA gene sequences of the isolated strains and related taxa was constructed using MEGA7 program [14] based on the neighbor-joining (NJ) algorithm [15].
성능/효과
The Micrococcaceae family isolates from Bgp40-b and Zdb130-b were identified as Pseudarthrobacter siccitolerans and the only Bacillales Family XII isolated from Bgb40-b was identified as Exiguobacterium mexicanum. In addition, B. altitudinis, B. cereus, B. licheniformis, B. mobilis, B. pacificus, B. paralicheniformis, B. siamensis, B. tequilensis, Lysinibacillus fusisormis, L. mangiferiumii, L. xylanilyticus, and P. siccitolerans were also isolated from the four bentonite samples. Microorganisms such as that from the Bacillaceae family can produce a variety of hydrolytic enzymes, including α-amylase, cellulase, and protease, which are potentially of great industrial value [20].
toyonensis enhanced their health and growth performances by adding to livestock feed [18]. In the Bdb130-1 sample, only three Paenibacillaceae were isolated, which were identified as Brevibacillus invocatus and Paenibacillus motobuensis. Interestingly, two isolated strains MBLB1234 and MBLB1235 clustered with P.
The 16S rRNA gene sequence of strain MBLB1234 was deposited in GenBank with accession number MG333459 and the further study on the polyphasic taxonomic analyses for this strain MBLB1234 will be conducted to report this strain to be novel Paenibacillus species. The Micrococcaceae family isolates from Bgp40-b and Zdb130-b were identified as Pseudarthrobacter siccitolerans and the only Bacillales Family XII isolated from Bgb40-b was identified as Exiguobacterium mexicanum. In addition, B.
후속연구
In addition, the isolated strains were revealed to present high hydrolytic enzyme activities for α-amylase, cellulase, and protease which could be useful for various industrial fields including food and pharmaceutical industries. Finally, this study could provide the basic microbial information in bentonite as one of clay minerals and industrial applications by bentonite-derived functional microorganisms.
65% of 16S rRNA gene sequence similarity could be as the threshold to determine a novel species [19], these two strains could be therefore considered new species within the genus Paenibacillus. The 16S rRNA gene sequence of strain MBLB1234 was deposited in GenBank with accession number MG333459 and the further study on the polyphasic taxonomic analyses for this strain MBLB1234 will be conducted to report this strain to be novel Paenibacillus species. The Micrococcaceae family isolates from Bgp40-b and Zdb130-b were identified as Pseudarthrobacter siccitolerans and the only Bacillales Family XII isolated from Bgb40-b was identified as Exiguobacterium mexicanum.
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