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.
참고문헌 (29)
Bergaya F, Lagaly G. 2006. General introduction: clays, clay minerals, and clay science. Develop. Clay Sci. 1: 1-18.
Seppala A, Puhakka E, Olin M. 2016. Effect of layer charge on the crystalline swelling of $Na^+$ , $K^+$ and $Ca^{2+}$ montmorillonites: DFT and molecular dynamics studies. Clay Miner. 51: 197-211.
Gonzalez-Pradas E, Villafranca-Sanchez M, Gallego-Campo A. 1993. Influence of the physical-chemistry properties of an acidactivated bentonite in the bleaching of olive oil. J. Chem. Technol. Biotechnol. 57: 213-216.
Allo WA, Murray HH. 2004. Mineralogy, chemistry and potential applications of a white bentonite in San Juan province, Argentina. Appl. Clay Sci. 25: 237-243.
Bereket G, Aroguz AZ, Ozel MZ. 1997. Removal of Pb(II), Cd(II), Cu(II), and Zn(II) from aqueous solutions by adsorption on bentonite. J. Colloid Interf. Sci. 187: 338-343.
Kang J, Chung WH, Nam YD, Kim D, Seo SM, Lim SL, et al. 2018. Impact of clay minerals on bacterial diversity during the fermentation process of kimchi. Appl. Clay Sci. 154: 64-72.
Seo SM, Kim D, Kim D, Kim JH, Lee YJ, Roh KM, et al. 2018. A simple synthesis of nitrate cancrinite from natural bentonite. J. Porous Mat. 25: 1561-1565 .
Roh SW, Sung Y, Nam YD, Chang HW, Kim KH, Yoon JH, et al. 2008. Arthrobacter soli sp. nov., a novel bacterium isolated from wastewater reservoir sediment. J. Microbiol. 46: 40-44.
Yoon SH, Ha SM, Kwon S, Lim J, Kim Y, Seo H, et al. 2017. Introducing EzBioCloud: a taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies. Int. J. Syst. Evol. Microbiol. 67: 1613-1617.
Kimura M. 1980. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J. Mol. Evol. 16: 111-120.
Saitou N, Nei M. 1987. The neighbour-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4: 406-425.
Miller RA, Beno SM, Kent DJ, Carroll LM, Martin NH, Boor KJ, et al. 2016. Bacillus wiedmannii sp. nov., a psychrotolerant and cytotoxic Bacillus cereus group species isolated from dairy foods and dairy environments. Int. J. Syst. Evol. Microbiol. 66: 4744-4753.
Miller RA, Jian J, Beno SM, Wiedmann M, Kovac J. 2018. Intra clade variability in toxin production and cytotoxicity of Bacillus cereus group type strains and dairy-associated isolates. Appl. Environ. Microbiol. 84: 2479-2517.
Kantas D, Papatsiros VG, Tassis PD, Giavasis I, Bouki P, Tzika ED. 2015. A feed additive containing Bacillus toyonensis ( $Toyocerin(^{(R)})$ ) protects against enteric pathogens in postweaning piglets. J. Appl. Microbiol. 118: 727-738.
Kim M, Oh HS, Park SC, Chun J. 2014. Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequence similarity for species demarcation of prokaryotes. Int. J. Syst. Evol. Microbiol. 64: 346-351.
Moreno ML, Piubeli F, Bonfa MRL, Garcia MT, Durrant LR, Mellado E. 2012. Analysis and characterization of cultivable extremophilic hydrolytic bacterial community in heavy-metal-contaminated soils from the Atacama Desert and their biotechnological potentials. J. Appl. Microbiol. 113: 550-559.
Hendricks CW, Doyle JD, Hugley B. 1995. New solid medium for enumerating cellulose-utilizing bacteria in soil. Appl. Environ. Microbiol. 61: 2016-2019.
Anson ML. 1938. The estimation of pepsin, trypsin, papain and cathepsin with hemoglobin. J. Gen. Physiol. 22: 79-89.
Das K, Doley R, Mukherjee AK. 2004. Purification and biochemical characterization of a thermostable, alkaliphilic, extracellular ${\alpha}$ -amylase from Bacillus subtilis DM-03, a strain isolated from the traditional fermented food of India. Biotechnol. Appl. Biochem. 40: 291-298.
Jeon HH, Jung JY, Chun BH, Kim MD, Baek SY, Moon JY, et al. 2016. Screening and characterization of potential Bacillus starter cultures for fermenting low-salt soybean paste (Doenjang). J. Microbiol. Biotechnol. 26: 666-674.
Hill J, Nelson E, Tilman D, Polasky S, Tiffany D. 2006. Environmental, economic, and energetic costs and benefits of biodiesel and ethanol biofuels. Proc. Natl. Acad. Sci. USA 314: 1598-1600.
Ram L, Kaur K, Sharma S. 2014. Screening isolation and characterization of cellulase producing micro-organisms from soil. Int. J. Pharm. Sci. Inv. 3: 12-18.
Soni SK. 2014. Production of xylanase-cellulase complex by Bacillus subtilis NS7 for the biodegradation of agro waste residues. Lignocellulose J. 1: 196-209.
Sumantha A, Larroch C, Pandey A. 2006. Microbiology and industrial biotechnology of food-grade proteases: a perspective. Food Technol. Biotechnol. 44: 211-220.
Ouattara HG, Reverchon S, Niamke SL, Nasser W. 2017. Regulation of the synthesis of pulp degrading enzymes in Bacillus isolated from cocoa fermentation. Food Microbiol. 63: 255-262.
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