• 한국물환경학회지
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• 제38권6호
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• pp.282-291
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• 2022

Since the efficiency of wastewater treatment using microalgae differs depending on the metabolic characteristics of the species, it is important to understand the characteristics of target algae prior to the application in wastewater treatment. In this study, for the application of Arthrospira platensis to wastewater treatment, which is a filamentous alkaliphilic cyanobacteria, basic species specificity was identified and the possibility of application to wastewater treatment was investigated. As a result of the species specificity investigation, the specific growth rate between pH 7.0 and 11.0 showed the highest value near pH 9 at 0.25/day. The reason for the relatively low growth(0.08/day) at pH 11 was thought to be the CA(carbonic anhydrase) enzyme that is involved in carbon fixation during photosynthesis has the highest activity at pH 8.0 to 9.0, and at pH 11, CA activity was relatively low. In addition, A. platensis showed optimal growth at 400 PPFD(photosynthetic photon flux density) and 30℃, and this means that cyanobacteria such as A. platensis have a larger number of PS-I(photosystem I) than that of PS-II(photosystem II). It was speculated that it was because higher light intensity and temperature were required to sufficiently generate electrons to transfer to PS-I. Regarding the applicability of A. platensis, it was suggested that if a system using the synergistic effect of co-culture of A. platensis and bacteria was developed, a more efficient system would be possible. And different from single cocci, filamentous A. platensis expected to have a positive impact on harvesting, which is very important in the latter part of the wastewater treatment process.

• Journal of Microbiology and Biotechnology
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• 제22권12호
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• pp.1636-1643
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• 2012

Enzymatic pre-bleaching by modification of pulp fibers with xylanases is an attractive approach to reduce the consumption of toxic bleaching chemicals in the paper industry. In this study, an alkaliphilic endoxylanase gene was isolated from metagenomic DNA of a structurally stable thermophilic lignocellulose-degrading microbial consortium using amplification with conserved glycosyl hydrolase family 10 primers and subsequent genome walking. The full-length xylanase showed 78% sequence identity to an endo-${\beta}$-1,4-xylanase of Clostridium phytofermentans and was expressed in a mature form with an N-terminal His6 tag fusion in Escherichia coli. The recombinant xylanase Xyn3F was thermotolerant and alkaliphilic, working optimally at $65-70^{\circ}C$ with an optimal pH at 9-10 and retaining >80% activity at pH 9, $60^{\circ}C$ for 1 h. Xyn3F showed a $V_{max}$ of 2,327 IU/mg and $K_m$ of 3.5 mg/ml on birchwood xylan. Pre-bleaching of industrial eucalyptus pulp with no prior pH adjustment (pH 9) using Xyn3F at 50 IU/g dried pulp led to 4.5-5.1% increase in final pulp brightness and 90.4-102.4% increase in whiteness after a single-step hypochlorite bleaching over the untreated pulp, which allowed at least 20% decrease in hypochlorite consumption to achieve the same final bleaching indices. The alkaliphilic xylanase is promising for application in an environmentally friendly bleaching step of kraft and soda pulps with no requirement for pH adjustment, leading to improved economic feasibility of the process.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2005년도 International Meeting of the Microbiological Society of Korea
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• pp.187.4-187
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• 2005

• Journal of Microbiology and Biotechnology
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• 제25권5호
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• pp.681-686
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• 2015

The purpose of this study was to find a cold-adapted and surfactant-stable alginate lyase as a candidate for biotechnological and industrial applications. The gene for a new alginate lyase, AlyL1, from Agarivorans sp. L11 was cloned and expressed in Escherichia coli. The recombinant AlyL1 was most active at 40℃ (1,370 U/mg). It was a cold-adapted alginate lyase, which showed 54.5% and 72.1% of maximum activity at 15℃ and 20℃, respectively. AlyL1 was an alkaliphilic enzyme and most active at pH 8.6. In addition, it showed high stability in the presence of various surfactants at a high concentration (from 0.1% to 1% (w/v)). AlyL1 was an endo-type alginate lyase that degraded both polyM and polyG blocks, yielding disaccharides and trisaccharides as the main products. This is the first report of the cloning and functional expression of a cold-adapted and surfactant-stable alginate lyase. AlyL1 might be an interesting candidate for biotechnological and industrial applications.

• Journal of Microbiology and Biotechnology
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• 제22권3호
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• pp.385-389
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• 2012

The application of microorganisms in the field of construction material is rapidly increasing worldwide; however, almost all studies that were investigated were bacterial sources with mineral-producing activity and not with organic substances. The difference in the efficiency of using bacteria as an organic agent is that it could improve the durability of cement material. This study aimed to assess the use of biofilm-forming microorganisms as binding agents to increase the compressive strength of cement-sand material. We isolated 13 alkaliphilic biofilmforming bacteria (ABB) from a cement tetrapod block in the West Sea, Korea. Using 16S RNA sequence analysis, the ABB were partially identified as Bacillus algicola KNUC501 and Exiguobacterium marinum KNUC513. KNUC513 was selected for further study following analysis of pH and biofilm formation. Cement-sand mortar cubes containing KNUC513 exhibited greater compressive strength than mineral-forming bacteria (Sporosarcina pasteurii and Arthrobacter crystallopoietes KNUC403). To determine the biofilm effect, Dnase I was used to suppress the biofilm formation of KNUC513. Field emission scanning electron microscopy image revealed the direct involvement of organic-inorganic substance in cement-sand mortar.

• 한국지반공학회논문집
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• 제30권1호
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• pp.93-101
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• 2014

본 논문에서는 자원 재활용을 위해 시멘트를 전혀 사용하지 않고 잠재 수경성을 지닌 고로슬래그와 알칼리 활성화제를 이용하여 모래를 고결시키는 연구를 수행하였다. 기존 수산화칼슘이나 수산화나트륨과 같은 화학적 알칼리 활성화제뿐 아니라 pH 10 이상에서 생존하는 극한미생물을 화학적 알칼리 활성화제에 혼합한 미생물 알칼리 활성화제를 개발하여 흙의 고결 가능성을 평가하였다. 낙동강모래에 고로슬래그의 함유량을 네 종류(4, 8, 12, 16%)로 달리하면서 화학적 또는 미생물 알칼리 활성화제를 혼합하여 공시체를 제작한 다음 7일 동안 대기중 양생시킨 후 일축압축시험을 실시하였다. 알칼리 활성화제의 종류에 관계없이 고로슬래그의 함유량이 4%에서 16%로 증가함에 따라 건조밀도가 증가하면서 일축압축강도는 평균 178kPa에서 2,435kPa까지 증가하였다. 화학적 알칼리 활성화제를 사용한 경우, 수산화칼슘이 포함된 공시체의 일축압축강도가 수산화나트륨을 사용한 경우보다 5-54% 정도 높게 나타났다. 한편 본 연구에서 개발한 미생물 알칼리 활성화제를 사용한 경우, 수산화칼슘 성분이 포함된 공시체의 경우에는 화학적 알칼리 활성화제보다 일축압축강도가 11-60% 감소하였으나, 수산화나트륨이 포함된 경우에는 일축압축강도가 19-121% 증가하였다. 고결된 공시체에서 C-S-H 화합물이 생성되었으며, SEM분석에서 고로슬래그 함유량이 증가할수록 수화물의 양도 증가하였다.

• 한국지반공학회논문집
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• 제31권1호
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• pp.37-46
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• 2015

본 연구에서는 고로슬래그의 수화반응을 일으키는 극한미생물(Bacillus halodurans) 알칼리 활성화제를 사용하여 현장 지반을 고결시키는 연구를 수행하였다. 현장 토사를 고결시키기 위해 저가의 미생물 배양액을 대량으로 제조하였으며, 제조된 미생물 배양액에 대한 생장실험을 실시하여 기존 미생물 배양액과의 효율성을 비교하였다. 현장 적용은 고로슬래그와 미생물 배양액을 혼합한 알칼리 활성화제로 고결된 지반(미생물 고결토), 보통 포틀랜드 시멘트로 고결된 지반(시멘트 고결토), 그리고 무처리된 지반(무처리토)으로 나누어 시험 시공하였다. 현장 지반 3곳 모두 동일한 크기인 가로 2.6m, 세로 4m, 깊이 0.2m로 시공하였다. 현장 시공 후 28일에 코어를 채취하여 일축압축시험을 실시하였으며, 무처리토 지반은 토베인시험으로 지반의 강도를 평가하였다. 본 연구에서 개발한 미생물 고결토는 시멘트 고결토에 비해 약 1/5 정도 낮은 강도를 보였으나, 무처리토에 비해서는 약 6배 정도 높은 강도를 발휘하였다. 또한, 미생물 고결토의 pH는 10으로 11 이상인 시멘트 고결토보다 낮아 상대적으로 친환경적인 것으로 판단되었으며, SEM-EDS 분석을 통하여 고결도와 고결 물질인 C-S-H 수화물이 생성됨을 확인할 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제16권8호
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• pp.1255-1261
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• 2006

An alkaliphilic bacterium, Bacillus sp. strain BK, was found to produce extracellular cellulase-free xylanolytic enzymes with xylan-binding activity. Since the pellet-bound xylanase is eluted with 2% TEA from the pellet of the culture, they contain a xylan-binding region that is stronger than the xylan-binding xylanase of the extracellular enzyme. The xylanases had a different molecular weight and xylan-binding ability. The enzyme activity of xylanase in the extracellular fraction was 6 times higher than in the pellet-bound enzyme. Among the enzymes, xylanase had the highest enzyme activity. When Bacillus sp. strain BK was grown in pH 10.5 alkaline medium containing xylan as the sole carbon source, the bacterium produced xylanase, arabinofuranosidase, acetyl esterase, and $\beta$-xylosidase with specific activities of 1.23, 0.11, 0.06, and 0.04 unit per mg of protein, respectively. However, there was no cellulase activity detected in the crude enzyme preparation. The hydrolysis of agricultural residues and kraft pulps by the xylanolytic enzymes was examined at 50$^{\circ}C$ and pH 7.0. The rate of xylan hydrolysis in com hull was higher than those of sugarcane bagasse, rice straw, com cop, rice husk, and rice bran. In contrast, the rate of xylan hydrolysis in sugarcane pulp was 2.01 and 3.52 times higher than those of eucalyptus and pine pulp, respectively. In conclusion, this enzyme can be used to hydrolyze xylan in agricultural residues and kraft pulps to breach and regenerate paper from recycled environmental resources.

• Journal of Microbiology and Biotechnology
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• 제20권4호
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• pp.670-677
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• 2010

An alkaline pectate lyase, Bsp165PelA, was purified to homogeneity from the culture broth of alkaliphilic Bacillus sp. N16-5. The enzyme showed a specific activity as high as 1,000 U/mg and had optimum activity at pH 11.5 and $50^{\circ}C$. It was composed of a single polypeptide chain with a molecular mass of 42 kDa deduced from SDS-PAGE, and its isoelectric point was around pH 6.0. It could efficiently depolymerize polygalacturonate and pectin. Characterization of product formation revealed unsaturated digalacturonate and trigalacturonate as the main products. The pectate lyase gene (pelA) contained an open reading frame (ORF) of 1,089 bp, encoding a 36-amino acids signal peptide and a mature protein of 326 amino acids with a calculated molecular mass of 35.943 Da. The deduced amino acid sequence from the pelA ORF exhibited significant homology to those of known pectate lyases in polysaccharide lyase family 1. Some conserved active-site amino acids were found in the deduced amino acid sequence of Bsp165PelA. $Ca^{2+}$ was not required for activity on pectic substrates.

• Journal of Microbiology and Biotechnology
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• 제18권12호
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• pp.1908-1914
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• 2008

An alkaliphilic bacterium, Bacillus clausii SKAL-16, was isolated from soil that had been contaminated with vegetable oil. The optimal pH and general pH range for bacterial growth was 8, and 7 to 10, respectively. The bacterium could grow on tributyrin and glycerol, but could not grow on acetate and butyrate. The SKAL-16 strain excreted butyric acid during growth on tributyrin, and selectively ingested glycerol during growth on a mixture of butyric acid and glycerol. The SKAL-16 generated intracellular lipase, but did not produce esterase and extracellular lipase. The DNA fragment amplified with the chromosomal DNA of SKAL-16 and primers designed on the basis of the esterase-coding gene of Bacillus clausii KSM-KI6 was not identical with the esterase-coding gene contained in the GenBank database. Pyruvate dehydrogenase, isocitrate dehydrogenase, and malate dehydrogenase activities were detected in the cell-free extract (crude enzyme).