Isolation,identification and desulfurization mechanism of a sulfur-oxidizing bacterium with salt tolerance北大核心CSCD

Micro-organism with efficient desulfurization performance is a key factor in the biological desulfurization technology. This study aimed to seek such a sulfur-oxidizing strain and understand its desulfurization mechanism. Wastewater in a sewage station of natural gas purification plant was used to screen the sulfide-oxidizing strain, and to identify it based on sequence similarity analysis of 16S rDNA and the morphological characteristics. Thiosulfate was used as substrate for investigating the sulfur oxidation performance and salinity tolerance; the OD600, content change of thiosulfate, sulfate, sulfur, pH and total alkalinity in the cultural system were also investigated. The strain DS-B was found to share the highest sequence similarity with Thioalkalivibrio thiocyanoxidans ARh2, therefore determined as Thioalkalivibrio. At the optimum temperature of 35 °C for growth and degradation, the removal efficiency of thiosulfate could reach 98.7% after 7 days. Strain DS-B had strong resistance to thiosulfate, and the optimal concentration of S2O32- was 2 × 104 mg/L. The analysis for sulfur oxides showed that it could oxidize thiosulfate by the pathway of S2O32-→SO42- / S2O32- → S → SO42-. Therefore the strain DS-B is a sulfur-oxidizing bacterium with great application prospect for its strong salt tolerance and conspicuous removal capability for thiosulfate.     

Isolation,identification, and the degradation characteristics of a thiobencarb-degrading bacterium Bacillus sp. T2北大核心CSCD

Thiobencarb, a thiocarbamate herbicide, is widely used to control weeds in rice paddies. Screening for highly efficient thiobencarb-degrading bacteria is important for the bioremediation of thiobencarb-contaminated environments. The aim of this study was to isolate and identify highly efficient thiobencarb-degrading bacteria and to identify the degradation pathway and the degrading properties. The thiobencarb-degrading strain was isolated using methods of microbiological acclimation and enrichment and was then identified using a 16S rRNA phylogenetic analysis. The degrading properties of the isolated bacterium were determined by single-factor experiments, and the degradation products were identified using gas chromatography-mass spectrometry (GC-MS). A thiobencarb-degrading strain T2, which can utilize thiobencarb as the sole source of carbon for energy and growth, was isolated from paddy soil. Strain T2 degraded more than 98.3% of 0.4 mmol/L of thiobencarb within 36 h. It was preliminarily identified as Bacillus sp. T2 according to the 16S rRNA gene analysis and from its morphological, physiological, and biochemical characteristics. The metabolic products of the thiobencarb degradation for strain T2 were identified as 4-chlorobenzyl mercaptan, 4-chlorobenzaldehyde, and 4-chlorobenzoic acid by the GC-MS. Based on metabolite identification, it was speculated that thiobencarb degradation in strain T2 was initiated by the hydrolysis of the thioester bond to produce 4-chlorobenzyl mercaptan, which was further oxidized to 4-chlorobenzaldehyde and 4-chlorobenzoic acid. The thiobencarb degradation that was initiated by the hydrolysis of the thioester bond by strain T2 is a new metabolic pathway, which provides valuable research material and reliable experimental data for revealing the metabolic process and mechanism of thiobencarb microbial degradation in soil. The strain Bacillus sp. T2 has a very high degradation efficiency, suggesting it is a good prospect for microbial remediation in thiobencarb-polluted environments. © 2018 Science Press. All rights reserved.     

Isolation,identification, and the degradation characteristics of a BDE-47 degrading strain北大核心CSCD

In an effort to remove BDE-47 residues from the environment, a bacterial strain that is capable of utilizing BDE-47 as the sole carbon source was isolated and screened from soil collected from an e-waste recycling area in Tianjin to analyze the degradation properties. The strain was preliminarily identified as Enterobacter sp. according to a 16S rDNA gene sequence analysis. The strain degraded 35.8% of 525 μg/L of BDE-47 in 35 d when the initial concentration of bacteria was 7.1 × 105 cells/ mL. The product of the biodegradation of BDE-47 was BDE-28. The biodegradation of BDE-47 fit well with first-order kinetics, and its degradation kinetics was ln Ct = - 0.104t + 6.22. With the addition of an electron acceptor, such as Fe3+, SO4 2- and NO3 -, the BDE-47 degradation rate was significantly increased to 49.8%, 59.1%, and 67.3%, respectively. The above results revealed that the strain could degrade BDE-47, which is of importance in the application of environmental bioremediation of BDE-47. © 2018 Science Press. All rights reserved.     

Screening of the marine actinomycetes with anti-complement activity and isolation of the active fractions北大核心CSCD

Studies on anti-complement agents are essential for the development of drugs to treat various diseases caused by excessive or abnormal activation of the complement system. However, studies on microbial-derived anti-complement agents are still very limited. The anti-complement activities of 42 marine-derived actinomycete strains isolated from the sediment samples collected from Xinghai Bay in Dalian were studied using the hemolysis method. In addition, the active fractions of the strain Streptomyces sp. DUT11 were separated by C18 and Sephadex LH-20 column chromatography, and the active compounds with anti-complement activity were identified. Streptomyces strains S187, M5, S088, M8, S063, DUT11 and MD16 showed good anti-complement activities upon being cultured in TSB medium, with the strain DUT11 showing the best anti-complement activity among these strains. When different fermentation media were tested, the highest anti-complement activity of DUT11 was observed in M33 fermentation medium. The anti-complement activity was 56.5% for the extract of supernatant and 60.8% for the extract of mycelia, respectively. Furthermore, three compounds with anti-complement activity were obtained from the active components, which were identified as 3-indolecarboxylic acid (1), p-hydroxy benzoic acid (2) and 2-pyrrolecarboxylic acid (3). These results demonstrated that marine-derived streptomycetes can be employed to produce active compounds with anti-complement activity. This study presents a new alternative for the utilization of marine actinomycetes and provides a basis for the exploration of new anti-complement agents. © 2018 Science Press. All rights reserved.     

Screening and characterization of extremophiles in spacecraft assembly environment北大核心CSCD

Spacecrafts need to strictly control microorganisms before entering space flight. The Spacecraft Assembly, Integration, and Test Center (AITC) is an important environmental source for spacecrafts to carry microorganisms. The assembly clean room has characteristics of ventilation, dryness, and lack of nutrients that are not conducive to the growth and reproduction of microorganisms, except for extremophiles. In this study, based on the internal air and surface environment of the AITC in China, 13 strains of bacteria were identified by plate culture and 16S sequence analysis, and their extreme environmental tolerance, antibiotic resistance, and film-forming ability were tested. All these bacteria belonged to the Firmicutes and Proteobacteria phyla, and nine strains belonged to the Bacillus genus. All 13 strains of bacteria were salt-tolerant, acid-tolerant, and alkali-tolerant, and 69.2% of the bacteria survived heat shock at 80 ℃. Among these, one strain of Sphingomonas sp. JCM7513 isolated from the surface environment was insensitive to all the tested antibiotics and had strong drug resistance, while the other 12 strains were sensitive to most β-lactam antibiotics but had strong resistance to tetracyclic antibiotics and erythromycin. Most of the isolated bacteria exhibited strong biofilm-forming abilities. The study showed that there are a certain number of extremophiles in the spacecraft assembly environment. To protect spacecrafts from biological corrosion and planetary protection forward pollution, more effective monitoring and elimination methods are needed. © 2022 Science Press. All rights reserved.     

Screening and degradation characteristics of a tetracycline-degrading bacterial strain北大核心CSCD

Tetracycline is widely used in livestock and poultry breeding industry, which can cause serious problems to the environment. Antibiotic pollution has become an important environmental issue. This study aimed to isolate and identify a well-functioning tetracycline-degrading bacteria strain from activated sludge and to investigate its optimum degradation conditions. The strain was identified through morphological features, Gram staining, and the sequence analysis of 16S rRNA. Furthermore, the temperature, initial pH of the medium, inoculation amount, and type of metallic salt were analyzed to investigate the tetracycline degradation performance of the isolated strain. Based on the single factor test, the method of response surface analysis was adopted to optimize the degradation condition. The strain was named TTC-1 and identified as Klebsiella pneumoniae. The optimum condition for tetracycline degradation was determined as follows: temperature of 34.4 °C, pH of 7.22, and MnSO4 concentration of 0.32 g/L. Under this optimum condition, the predicted tetracycline degradation rate was 93.77%, whereas the observed value was 94.26%. The experimental results showed that the proposed model had high accuracy. TTC-1 showed a good performance in degrading tetracycline, which can provide reference for the bacteria during the biological treatment of tetracycline containing wastewater. © 2018 Science Press. All rights reserved.     

The oligopeptides production by the feather-degrading bacterium Bacillus methylotrophicus H0北大核心CSCD

Due to the high nutritive value of oligopeptides and the waste of feather resources, this study aimed at screening efficient strains of bacteria able to rapidly degrade feathers and produce large quantities of value-added oligopeptides. In order to assess the potential yield of oligopeptides, the promising strain H0 was selected from 16 feather-degrading microorganisms. To identify the strain, we analyzed the morphological and physiological characteristics of different strains, and carried out a gene sequence analysis of their 16S rRNAs. A single factor experiment was used to promote feather degradation and oligopeptide production, and the characteristics of the oligopeptides produced were also analyzed by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS). The strain was identified as Bacillus methylotrophicus. The optimal initial pH and temperature for oligopeptide production were 11 and 40 °C, respectively. After 72 h of fermentation under these optimal conditions, the feathers were almost completely degraded, with a 38.19% of oligopeptides (accounting for 67.53% of the total soluble peptides) and a 11.11% of free amino acids produced. LC-MS/MS analysis indicated that the oligopeptides were mainly short peptides containing 5-10 amino acids, with a molecular mass (Mr) of less than 1 300. Moreover, the peptides were abundant in branched-chain amino acids, that might be responsible for the antioxidant property of the feather hydrolysate. Our results demonstrate the great capability of B. methylotrophicus H0 in feather degradation and oligopeptide production. This research provides a high-quality microorganism resource, and the scientific basis for the development of feather-derived oligopeptide products. © 2018 Science Press. All rights reserved.     

Genome-wide identification and hormone expression patterns of the MaPIF family of banana genes北大核心CSCD

植物光敏色素作用因子(phytochrome interacting factor,PIF)是广泛分布于植物体内的一种转录因子,在植物的生长发育方面有着重要的作用.基于香蕉基因组数据,对香蕉MaPIF基因家族进行基因组鉴定,采用生物信息学分析方法对其进行命名,分析理化性质、蛋白质结构、基因结构、启动子顺势作用元件以及构建系统进化树;分析PIF家族在不同激素处理下的表达情况.结果显示,香蕉MaPIF家族有7个成员,均含有高度保守的bHLH结构域;编码区长度在1 116-2001bp之间,至少包含5个内含子,且大部分位于细胞外;进化树结果可以发现与拟南芥、水稻以及玉米PIF的亲缘关系较近;顺式作用元件预测结果显示,MaPIF上存在多种与激素和光相关的响应元件.qRT-PCR结果显示,MaPIF3-1、MaPIF4、MaPIF4-1在生长素(IAA)、赤霉素(GA)、生长素抑制剂(NPA)处理中均有显著表达,除此之外,所有成员在脱落酸(ABA)处理下均有明显表达.本研究表明MaPIF在香蕉生长发育中激素调控有重要作用.(图7表3参45)     

Isolation,identification, and biological characteristics of a wild Chaga mushroom北大核心CSCD

In this study, a pure culture strain (numbered as F1501) was obtained using tissue separation and purification methods from the sclerotia of Chaga mushroom from Russia. Further researches included studies on classification, optimum growth conditions of mycelia, extracellular polysaccharides from fermentation broth and their antioxidant activity, and artificial acclimation. According to the identification of the internal transcribed spacer (ITS) region, it was confirmed that F1501 was a species of the genus Inonotus and family Hymenochaetaceae, which had 99% similarity with Inonotus obliquus. F1501 was further classified as I. obliquus based on the morphological characteristics of its mycelia and sclerotia. The optimal carbon source, nitrogen source, C/N ratio, growth factor, temperature, and pH value for the growth of the F1501 strain mycelia were maltose, beef extracts, 10/1, B2, 28 °C, and 8.0, respectively. Liquid fermentation of F1501 was performed using PD media and a 10% inoculation amount at 28 °C and 150 r/min for 7 d. The content of polysaccharides in the fermentation broth was 476.32 mg/L, with a total antioxidant activity of 0.19 mmol/L (Trolox) and hydroxyl free radical-scavenging activity of 72.7%. Artificial acclimation study revealed that the fruiting body-like structure was obtained using cottonseed hulls as the main substrate. In the present study, a new strain of I. obliquus and its biological characteristics were explored, which could provide a theoretical basis for the artificial acclimation and development of wild mushrooms. © 2018 Science Press. All rights reserved.     

Protoplast preparation and regeneration conditions of Bacillus subtilis ZGL14 producing alkaline pectinase北大核心CSCD

The protoplast fusion technique is one of the important microbial breeding methods. The main factors that affect protoplast preparation and regeneration were investigated using Bacillus subtilis ZGL14 as the starter strain. The optimal conditions for protoplast preparation and regeneration were determined in this study by optimizing the cell age, lysozyme concentration, enzymolysis time, enzymolysis temperature, pH values of the osmotic pressure stabilizer, regeneration medium types, and culture type. The results indicated that the optimal condition for protoplast preparation of B. subtilis ZGL14 was as follows: 9 h of cell age, 0.2 mg/mL of lysozyme concentration, 10 min of enzymolysis time, 32 °C of enzymolysis temperature, and 7.0 of osmotic pressure stabilizer pH value. Under this condition, the protoplast formation and regeneration rates were > 98% and > 30%, respectively. A good protoplast regeneration effect can be achieved using the regeneration medium with sodium succinate as osmotic pressure stabilizer in combination with Ca2+. The single-layer culture was better than double-layer for regeneration. The optimal condition for protoplast preparation and regeneration of B. subtilis ZGL14 was determined in this study, which provides technical support for protoplast fusion breeding of B. subtilis. © 2018 Science Press. All rights reserved.     

Isolation,identification, and the biodegradation characteristics of a benzoylurea insecticides-degrading strain北大核心CSCD

A chlorbenzuron, diflubenzuron, and hexaflumuron-degrading bacterium strain M6, was isolated from the activated sludge of an insecticide factory. The strain was identified as Achromobacter sp. according to an analysis on the 16S rRNA gene sequences, morphological, and physiological characteristics. Strain M6 could degrade more than 91% of 100 mg/L chlorbenzuron, diflubenzuron, and hexaflumuron within 48 hours, which could act as the sole carbon source. Strain M6 showed more chlorbenzuron degradation at a temperature range between 25 and 40 ℃ and a pH range between 6.0 and 8.0. The optimal temperature and the initial pH of medium for chlorbenzuron degradation by strain M6 were 30 ℃ and 7.0, respectively; the maximum chlorbenzuron tolerated concentration of strain M6 was as high as 400 mg/L. Strain M6 hydrolyzed 4-acetaminophenol into a purple-red product. Moreover, an approximately 1.4 kb DNA fragment, which could be expressed into an amidase to degrade amide pesticides, was amplified from the genomic DNA of strain M6. The results preliminarily proved that 3 benzoylurea insecticides could be degraded because of strain M6 hydrolyzing their amide bonds. This study obtained a highly efficient degrading strain and provided new resources and valuable information on benzoylurea insecticide degradation. © 2018 Science Press. All rights reserved.     

Degradation of chitin and antagonism in Phomopsis asparagi by Bacillus amyloliquefaciens MY001北大核心CSCD

About 10 billion tons of chitin, a natural polymer, is generated annually. However, the utilization of chitin was hindered because of the lack of a clean and effective degradation process. In this study, bacterial strains with chitosanase-producing ability were isolated from soil and identified using 16S rDNA, gyrA, and gyrB gene sequence analysis. In addition, the degradation of insoluble chitin and antifungal capability of the bacterial strains were investigated. One selected isolate was identified as Bacillus amyloliquefaciens strain MY001. It showed antagonistic effect on Phomopsis asparagi (Sacc.) Bubak and could degrade insoluble chitin. The combination of chitin/chitosan and MY001 strain could enhance the inhibition on the growth of P. asparagi. In conclusion, strain B. amyloliquefaciens MY001 can directly degrade chitin. The combination of MY001 strain with chitin shows more effective inhibition on P. asparagi growth. These results suggest the direct utilization of chitin against fungal diseases of crops. © 2018 Science Press. All rights reserved.     

Screening and identification of bacterial strain producing (R)-1-phenyl-1,2-Ethanediol by efficient whole-cell catalytic reduction system北大核心CSCD

(R)-1-Phenyl-1,2-ethanediol [(R)-PED] is an important chiral intermediate used to synthesize pharmaceutical intermediates. The production of (R)-PED is of great practical significance. A novel bacterial strain HBU-SI7 isolated from soil samples was screened using a-hydroxyacetophenone (2-HAP) as substrate, which is capable of enantioselective reduction of 2-HAP to (R)-PED. HBU-SI7 was identified as Rhodococcus sp. by bacterial morphological observation and 16S rDNA analysis. The analysis of whole-cell catalytic system showed that the bioconversion of 2-HAP reached 96.2% in the two-phase reaction system (dibutyl phthalate/phosphate buffer = 1/3, V/V) with the substrate concentration of 3.0 g/L, and 99.3% of e.e. for (R)-PED was achieved. A new enantio selective reduction system with the whole cells of Rhodococcus sp. was established for efficient production of (R)-PED, which has potential research and application values. © 2018 Science Press. All rights reserved.     

Isolation,identification, culture conditions,and laccase production of white rot fungus北大核心CSCD

The white rot fungi are members of Basidiomycota, which can degrade lignin and form white rot. They are high producers of extracellular laccases. In the present study, pure culture strain of high-temperature and high-laccase production types (numbered as BUA-01) was isolated from the fruiting bodies of a white rot fungus collected in the campus of Beijing University of Agriculture. The taxonomic characteristic was determined based on morphological and ITS sequence analysis. Furthermore, the optimal culture conditions for the mycelia were determined, including carbon source, nitrogen source, C/N ratio, growth factors, temperature, and pH. Extracellular laccase production was investigated in liquid fermentation with different concentrations of Cu (CuSO4) as inducer. Decolorizing activity of the fermentation broth was assayed using three azo dyes: Evans blue, methyl orange, and eriochrome black T. The results showed that the strain possessed the highest homology toward Trametes hirsuta, with the homology ratio of 100% and the genetic distance of 0, suggesting that the strain BUA-01 belonged to the genus Trametes. The culture condition investigated revealed that the optimal condition for mycelia growth included the following: carbon source, starch; nitrogen source, soybean powder and yeast extract; C/N ratio, 40/1 and 10/1; temperature, 37 °C; and pH, 6.0-7.0. The assayed growth factors had no significant effect on mycelial growth. It demonstrated high laccase activity in liquid fermentation. The highest extracellular laccase activity of 1 081.33 ± 6.3 U/mL was observed in the broth with a Cu adjunction concentration of 0.25 mmol/L after a 96-h culture period. It was about 26-fold higher than that of the control group. The isolated strain exhibited significant decolorizing activity toward the azo dyes Evans blue, methyl orange, and eriochrome black T, with the decolorization rate at 12 h of 93.31% ± 0.16%, 92.37% ± 0.42%, 79.25% ± 0.64%, respectively. This suggests that the strain possesses potential applications in laccase production and dye degradation. © 2018 Science Press. All rights reserved.     

Cloning,expression, and functional identification of CaSWEET7 and CaSWEET15 genes in Canarium album北大核心CSCD

糖外排转运蛋白(sugars will eventually be exported transporters,SWEET)介导植物光合同化产物蔗糖的跨膜运输.以橄榄(Canarium album(Lour.)Raeusch.)果实为材料,通过气相色谱串联质谱(GC-MS)检测橄榄果实中糖组分及含量变化,利用RT-PCR技术克隆得到CaSWEET7和CaSWEET15基因开放阅读框(ORF)序列.结果表明:CaSWEET7和CaSWEET15基因ORF全长分别为774 bp和951 bp,分别编码长度为257个和316个氨基酸残基,具有2个MtN3_slv结构域;进化树分析表明,CaSWEET7属于CladeⅡ,CaSWEET15属于CladeⅢ.实时荧光定量PCR(qRT-PCR)技术检测其在不同发育时期的果实中相对表达量变化,结果表明,随着果实发育,CaSWEET7和CaSWEET15表达量逐渐递增,且与果实发育蔗糖含量变化呈显著正相关(相关性系数分别为0.931和0.904,P<0.05);利用酵母功能互补证明CaSWEET7和CaSWEET15基因编码的蛋白具有转运蔗糖能力.本研究表明CaSWEET7和CaSWEET15基因可能在橄榄果实蔗糖积累中发挥作用.(图9表2参44)