Isolation and Dye Decolorization of a Bacillus subtilis Strain LS02 Exhibiting Laccase Activity

2011 ◽  
Vol 183-185 ◽  
pp. 839-843
Author(s):  
Mei Hui Du ◽  
Min Zhao ◽  
Lei Lu ◽  
Tian Nv Wang ◽  
Tai Lun Li ◽  
...  

A newly isolated strain LS02 was estimated for its ability in dye decolorization. The LS02 strain was identified as Bacillus subtilis by the combination of physicochemical tests and 16S rDNA sequence analysis. The isolated strain could oxidize the laccase substrate syringaldazine, indicating the existence of laccase activity. B. subtilis LS02 grown well in the pH range of 5.0~9.0, and showed an optimum growth temperature at 37°C. Indigo carmine could be completely decolorized by B. subtilis LS02 after 4 days, whereas Remazol Brilliant Blue R, reactive black 5 and crystal violet were poorly decolorized. The result indicated that the laccase of B. subtilis LS02 may be suitable for the application in textile bleaching of indigo carmine.

2010 ◽  
Vol 113-116 ◽  
pp. 226-230 ◽  
Author(s):  
Chun Lei Wang ◽  
Min Zhao ◽  
Xing Dong Wei ◽  
Tai Lun Li ◽  
Lei Lu

Treatment of xenobiotic compounds such as textile dyes with bacterial laccases is limited to the acid pH range and moderate temperatures. A bacterial strain, designated as WD23, was isolated from forest soil using Luria-Bertani medium supplemented with 0.4 mmol/L Cu2+. The isolated strain was identified as Bacillus subtilis by physiological and biochemical tests and 16S rDNA sequence analysis. Here we charactered the spore-bound laccase of B. subtilis WD23 and used the laccase to decolorize dyes. The spores of the strain showed laccase-like activity, oxidizing syringaldazine, 2,6-dimethoxyphenol and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonate acid)(ABTS). The optimum pH and temperature for the spore-bound laccase were 6.8 and 60°C, respectively. It also showed higher stabilities over a broad pH range, the pH half-life was more than 6 months at pH 6.8. The spore laccase could efficiently decolorize 50~90% of anthraquinone and azo dyes in 24 h. The spore laccase can play an important role in bioremediation.


2010 ◽  
Vol 113-116 ◽  
pp. 725-729 ◽  
Author(s):  
Chun Lei Wang ◽  
Min Zhao ◽  
De Bin Li ◽  
Dai Zong Cui ◽  
Hong Yi Yang ◽  
...  

The strain Bacillus sp. WD23 exhibiting laccase activity was screened from forest soil. The M9 medium containing Cu2+ was used for enriching and isolating bacterial strains capable of oxidizing syringaldazine (SGZ). One isolated strain was identified as Bacillus subtilis WD23 based on the results of physiological and biochemical tests and 16S rDNA sequence analysis. The strain WD23 could grow at temperatures ranging from 20 to 55°C and showed optimum growth temperature and pH at 25°C and 7.0, respectively. The sporulation rate of the strain clearly correlated well with the laccase activity. The temperature half-life of the spore laccase was 2.5 h at 80°C and the pH half-life was 15 d at pH 9.0. Its spore laccase could decolorized 50~90% of Remazol brilliant blue R (RBBR), alizarin red, congo red, methyl orange and methyl violet, which suggests the potential application of spore laccase in dyestuff treatment.


1960 ◽  
Vol 27 (2) ◽  
pp. 221-234 ◽  
Author(s):  
Constance Higginbottom ◽  
Margaret M. Taylor

SummaryThe sterilization of homogenized milk at 115·5°C for 15 min in bottles having a partial vacuum in the headspace produced conditions inhibitory to the growth from very small numbers of spores ofBacillus subtilis, B. licheniformis, B. cereusandB. breviswhen compared with growth in the same milk sterilized in open bottles.B. circulansdiffered from the other strains tested in showing greater inhibition in milk sterilized in open bottles than in milk sterilized under partial vacuum.The extent of the inhibition became less as the size of the inoculum was increased. It became less also as the temperature of incubation approached the optimum growth temperature of the bacillus, and was influenced by the strain of the bacillus and the source of the milk but not by the degree of heat treatment within the range 107–117·5°C for 15 min. Inhibition was manifested by a prolongation of the lag phase, and in addition with some strains inhibition of spore germination could be demonstrated.Spore formation following vegetative growth occurred more readily in milk sterilized in open than in evacuated bottles.Milks sterilized under partial vacuum frequently failed to show any growth from small inocula in 30 days at 22°C although growth occurred readily in milk sterilized in open bottles.


2011 ◽  
Vol 183-185 ◽  
pp. 773-777 ◽  
Author(s):  
Jun Bo Pan ◽  
Min Zhao ◽  
Lei Lu ◽  
Mei Hui Du ◽  
Guo Fu Li ◽  
...  

Bacterial strains exhibiting laccase activity were isolated from the forest soil. A strain LC02 with syringaldazine oxidation ability was obtained using enrichment medium supplemented with copper ions. The isolated strain was identified as Bacillus amyloliquefaciens using physiological and biochemical tests as well as 16S rDNA sequence analysis. The characterization of spore laccase activity was investigated. The result showed that the optimum pH and temperature of the enzyme was 6.6 and 70°C, respectively. A great thermostability was observed for the spore laccase at 70°C. Laccase activity was strongly inhibited by 0.1 mmol/L NaN3, dithiothreitol and cysteine.


2012 ◽  
Vol 450-451 ◽  
pp. 573-581
Author(s):  
Yang Li ◽  
Cheng Gang Zheng

Three bacterial strains were isolated from oil contaminated soil samples in Yumen oilfield of China and selected due to their capacity of growing under extreme conditions and production of bioemulsifier. The isolates were identified as Brevibacillus sp. XS1, Geobacillus sp. XS2 and Geobacillus sp. XS3 according to 16S rDNA sequence and physiological methods, respectively. The isolates XS1, XS2 and XS3 were thermotolerant with the optimum growth temperature of 50 °C, 60 °C and 55 °C, respectively. All the three isolates were able to produce bioemulsifiers which had little effect on surface tension reduction. The bioemusifiers produced by the three isolates were purified with a production yield of 2.98g/L, 4.24g/L and 3.82 g/L, respectively. The bioemulsifiers were identified as anionic heteropolysaccharides by FTIR analysis and their average molecular weight and polydispersity were investigated by GPC method. The bioemulsifiers produced by Geobacillus sp. XS2 and XS3 exhibited a perfect stability over various temperature (up to 100 °C), pH (range from 2.0 to 12.0) and salinity (up to 30.0% of NaCl concentration) and were considered to be ideal candidates in MEOR process.


2017 ◽  
Vol 79 (7) ◽  
Author(s):  
Ameerah Tharek ◽  
Bay Hui Han ◽  
Shaza Eva Mohamad ◽  
Zaharah Ibrahim

The discharge of highly coloured azo dyes effluent has caused serious environmental damages. In this study, bacteria isolated from Antarctica seawater were screened for their ability to decolourise azo dye Reactive Black 5 (RB5). The selected bacterium was further investigated to study its ability to decolourise RB5. The best bacteria from Antarctica seawater that had the ability to decolourise RB5 was identified using 16S rDNA sequence analysis and revealed that the bacteria 15C shared 99% homology to Klebsiella pneumoniae. Selection of the most effective bacteria was followed by its acclimatisation to decolourise higher concentrations of RB5 by growing it in successively higher concentrations of RB5. Following that, optimization of RB5 decolourisation by the selected bacteria was performed using one factor at time (OFAT) including concentration of dye, pH and temperature. The results obtained indicated the optimal condition for decolourisation of RB5 using this bacterium was at pH 10 and 37°C in 70 mg/L RB5 with 98% decolourisation within 24 h under facultative anaerobic treatment. Besides that, 70% of COD removal was achieved after 96 h of sequential anaerobic and aerobic treatment of RB5. In addition, FTIR and HPLC were used to analyze the metabolite of RB5 decolourisation. Products of RB5 decolourisation was confirmed by the presence of sulphanilic acid in HPLC analyses and the changes observed in the functional groups of the FTIR spectrum suggesting the possibility of RB5 degradation. Ammonia test and carbon dioxide test showed higher concentration of ammonia and carbon dioxide that indicates the mineralisation of product after treatment.


2021 ◽  
Vol 9 ◽  
Author(s):  
Jing Yang ◽  
Jian Zhao ◽  
Jianchun Jiang ◽  
Hao Xu ◽  
Ning Zhang ◽  
...  

Alkali lignin-degrading Bacillus were isolated from forest soils in China and were identified as Bacillus subtilis TR-03 and Bacillus cereus TR-25 by 16S rDNA sequence analysis. Wherein TR-03 displayed optimal 26.72% alkali lignin (2 g/L) degradation at 7 days and 71.23% of Azure-B (0.01%) decolorization at 36 h of cultivation at 37°C. Ligninolytic enzyme analysis revealed that TR-03 was capable of depolymerizing alkali lignin effectively by the producing of lignin peroxidase and laccase, wherein higher laccase activity was cell-associated. At last, the physical and chemical changes of lignin via SEM and FTIR analysis was further observed to prove the lignin degradation by Bacillus subtilis TR-03.


2020 ◽  
Vol 12 (2) ◽  
pp. 162-168
Author(s):  
Thi Quyen Ha ◽  
Thi Thu Ha CHU

The nitrogen fixing bacteria (NFB) and phosphate solubilizing bacteria (PSB) are used widely for producing of microbiological fertilizers. This study aims to seek nitrogen-fixing and phosphate-solubilizing bacteria strains to add to the collection of candidate strains for producing single and multi-function microbiological fertilizers. From 40 soil samples of 8 fields for cultivating rice, medicinal plants and vegetables, 15 NFB strains and 12 PSB strains were isolated and determined the ability of fixing nitrogen and solubilizing inorganic phosphate compound through creation of NH4+ and PO4- in culture medium. Among 15 NFB strains, the fixing nitrogen activities of 7 strains were much higher than the remaining strains, including NFBR3, NFBV2, NFBM5, NFBM3, NFBM1, NFBV5 and NFBR2 with NH4+ concentration 18.85 mg/l, 18.41 mg/l, 17.32 mg/l, 16.19 mg/l, 15.49 mg/l, 12.83 mg/l and 12.57 mg/l, respectively Among 12 PSB strains, The ability of solubilizing phosphate of 5 strains were higher than the others, including PSBM2, PSBR1, PSBV1, PSBR5 and PSBR3 with PO4- concentration 14.49 mg/l, 11.83 mg/l, 11.33 mg/l, 10.65 mg/l, 10.37 mg/l, respectively. 3 NFB strains (NFBR3, NFBV2, NFBM5) and 3 PSB strains (PSBM2, PSBR1, PSBV1) with higher activity were identified by 16S-rDNA sequence analysis and comparing to some homologous sequences in genbank. The results showed that NFBR3 was identified as Azotobacter vinelandii, NFBV2 as Azopirillum brasilense, NFBM5 as Azotobacter chroococum, PSBM2 and PSBV1 as Pseudomonas fluorescens and PSBR1 as Bacillus subtilis. Vi khuẩn cố định nitơ (NFB) và vi khuẩn phân giải phosphate (PSB) được sử dụng rộng rãi trong sản xuất phân bón vi sinh. Nghiên cứu này nhằm mục đích tìm kiếm các chủng vi khuẩn cố định nitơ và hòa tan phosphate, bổ sung vào bộ sưu tập các chủng dự tuyển cho sản xuất phân bón vi sinh đơn và đa chức năng. Từ 40 mẫu đất của 8 ruộng trồng lúa, cây dược liệu và rau màu, 15 chủng NFB và 12 chủng PSB đã được phân lập và xác định khả năng cố định nitơ và phân giải phosphate vô cơ thông qua sự tạo thành NH4+ và PO4- trong môi trường nuôi cấy. Trong số 15 chủng NFB, có 7 chủng có hoạt tính cố định nitơ cao hơn những chủng còn lại, bao gồm các chủng NFBR3, NFBV2, NFBM5, NFBM3, NFBM1, NFBV5 và NFBR2 với nồng độ NH4+ lần lượt là 18.85mg/l, 18.41 mg/l, 17.32 mg/l, 16.19 mg/l, 15.49 mg/l, 12.83 mg/l và 12.5 7mg/l. Trong số 12, có 5 chủng có khả năng phân giải phosphate cao hơn những chủng khác, bao gồm chủng PSBM2, PSBR1, PSBV1, PSBR5 và PSBR3 với nồng độ PO4- lần lượt là 14.49 mg/l, 11.83 mg/l, 11.33 mg/l, 10.65 mg/l và 10.37 mg/l. Các chủng NFB và PSB này đều xuất hiện ơ các mẫu đất trồng lúa, đất trồng cây dược liệu và đất trồng rau màu. 3 chủng NFB và 3 chủng PSB với hoạt tính cố định nitơ và phân giải phosphate cao hơn được định loại bằng phân tích trình tự gen 16S-rDNA và so sánh với một số trình tự tương đồng trong genebank. Kết quả chỉ ra rằng chủng NFBR3 được định danh là Azotobacter vinelandii, chủng NFBV2 là Azopirillum brasilense, chủng NFBM5 là Azotobacter chroococum, chủng PSBM2 và chủng PSBV1 là Pseudomonas fluorescens và chủng PSBR1 là Bacillus subtilis.


2010 ◽  
Vol 60 (11) ◽  
pp. 2535-2539 ◽  
Author(s):  
Hui-Rong Li ◽  
Yong Yu ◽  
Wei Luo ◽  
Yin-Xin Zeng

Strain ZS314T was isolated from a sandy intertidal sediment sample collected from the coastal area off the Chinese Antarctic Zhongshan Station, east Antarctica (6 ° 22′ 13″ S 7 ° 21′ 41″ E). The cells were Gram-positive, motile, short rods. The temperature range for growth was 0–26 °C and the pH for growth ranged from 5 to 10, with optimum growth occurring within the temperature range 18–23 °C and pH range 6.0–8.0. Growth occurred in the presence of 0–6 % (w/v) NaCl, with optimum growth occurring in the presence of 2–4 % (w/v) NaCl. Strain ZS314T had MK-10 as the major menaquinone and anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0 as major fatty acids. The cell-wall peptidoglycan type was B2β with ornithine as the diagnostic diamino acid. The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The genomic DNA G+C content was approximately 67 mol%. Phylogenetic analysis based on 16S rRNA gene sequence similarity showed that strain ZS314T represents a new lineage in the family Microbacteriaceae. On the basis of the phylogenetic analyses and phenotypic characteristics, a new genus, namely Marisediminicola gen. nov., is proposed, harbouring the novel species Marisediminicola antarctica sp. nov. with the type strain ZS314T (=DSM 22350T =CCTCC AB 209077T).


2021 ◽  
Vol 67 (1) ◽  
Author(s):  
Mariko Takano ◽  
Masaya Nakamura ◽  
Masanobu Tabata

AbstractWe performed an analysis using isoelectric focusing to comprehensively clarify the isozyme composition of laccase derived from Japanese lacquer tree, Toxicodendron vernicifluum. When water extracts of acetone powder obtained from lacquer were subjected to isoelectric focusing, five bands within pI 7.35–9.30 and nine bands within pI 3.50–5.25 were detected using Coomassie staining. Similarly, laccase activity staining using guaiacol showed five bands within pI 7.35–9.30 and three bands within pI 3.50–4.25. However, laccase activity staining using gallic acid showed remarkable staining within pI 3.50–5.85, whereas staining was very weak within pI 7.35–9.30. When the water extracts of acetone powder were fractionated into the fractions containing bands within pI 7.35–9.30 and pI 3.50–5.85 by SP-Sepharose column chromatography, the former had a blue color and the latter a yellow color. The laccase activity was measured for each of the fractions in buffer solution in the pH range of 2.5–8.0. When syringaldazine, guaiacol, and 2,6-dimethoxyphenol were used as substrates, the yellow fraction showed considerably higher activity than the blue fraction for pH 5.5–7.5. When 3-methylcatechol and 4-methylcatechol were used as substrates, the yellow fraction showed higher activity for pH 4.5–6.5, and the blue fraction showed higher activity for pH 7.0–8.0. When 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) was used as the substrate, both fractions showed maximum activity at optimum pH of 3.0–4.0. Conventionally, in research on blue laccase derived from lacquer, the non-blue fraction corresponding to the yellow fraction lower than pI 6 has been removed during the purification process and thus has not been analyzed. Our results indicated that yellow laccase was present in the non-blue components of lacquer and that it may play a role in urushiol polymerization with previously reported blue laccase.


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