Studies on culture condition of new marine bacterium sp. SY01

New marine bacterium Zooshikella sp. SY01, producer of prodigiosin, was isolated from the seawaters of Sanya Bay. The culture conditions of this bacterium were investigated. Zooshikella sp. SY01 was cultured in 2216E media which contained tryptophan, histidine, lactonic acid, camphor, limonene, casein, diphenyl guanidine, coumarin and 1,3-dinitrobenzene, respectively. After 5 days cultivation, the extracts of different culture broths were detected by direct infusion mass spectroscopy using positive ESI mode. As the results, tryptophan, histidine and casein didn’t show any observable influences on the biosynthesis of prodigiosin. Lactonic acid, camphor, limonene, diphenyl guanidine, coumarin could inhibit the bacterium growth and prodigiosin biosynthesis to a certain extent, slower the culture broth to turn red. However, 1, 3-dinitrobenzene inhibited the bacteria to produce prodigiosin completely. MS data suggested that various metabolites with chemodiversity were produced in different culture media. In particular, a series of high-molecular-weight compounds with high relative abundances were observed in the medium containing limonene. To further optimize the culture condition, more new prodigiosin analogues and lead compounds can be obtained and the goal of “one strain-many compounds” can be achieved.     

Studies on culture condition of new marine bacterium Zooshikella sp. SY01

New marine bacterium Zooshikella sp. SY01, producer of prodigiosin, was isolated from the seawaters of Sanya Bay. The culture conditions of this bacterium were investigated. Zooshikella sp. SY01 was cultured in 2216E media which contained tryptophan, histidine, lactonic acid, camphor, limonene, casein, diphenyl guanidine, coumarin and 1,3-dinitrobenzene, respectively. After 5 days cultivation, the extracts of different culture broths were detected by direct infusion mass spectroscopy using positive ESI mode. As the results, tryptophan, histidine and casein didn’t show any observable influences on the biosynthesis of prodigiosin. Lactonic acid, camphor, limonene, diphenyl guanidine, coumarin could inhibit the bacterium growth and prodigiosin biosynthesis to a certain extent, slower the culture broth to turn red. However, 1,3-dinitrobenzene inhibited the bacteria to produce prodigiosin completely. MS data suggested that various metabolites with chemodiversity were produced in different culture media. In particular, a series of high-molecular-weight compounds with high relative abundances were observed in the medium containing limonene. To further optimize the culture condition, more new prodigiosin analogues and lead compounds can be obtained and the goal of “one strain-many compounds” can be achieved. __________ Translated from Acta Scientiarum Naturalium Universitatis Sunyatseni, 2007, 46(6): 55–58 [译自: 中山大学学报(自然科学版)]     

鞘氨醇单胞菌DP58降解吩嗪-1-羧酸的特性研究

考察了鞘氨醇单胞菌DP58对吩嗪-1-羧酸(phenazine-1-carboxylic acid, PCA)的降解特性.以PCA为唯一碳氮源配成无机盐培养基,通过改变PCA的初始浓度和不同的细菌接种量,发现PCA被降解时减少的浓度与细菌细胞数量的增量呈线性关系.通过正交试验设计,以不同温度、pH值、摇床转速的组合确定影响该菌株降解PCA的主要因素为pH值,其次是温度.为分析PCA基团结构与降解的关系,研究了该菌株对PCA的结构类似物吩嗪的降解,结果表明鞘氨醇单胞菌DP58降解吩嗪的能力有限. 这些实验结果对今后进一步研究降解途径,从而采取措施抑制PCA降解,有十分重要的意义.     

鞘氨醇单胞菌脱硫丁苯橡胶胶粉填充丁苯橡胶性能的研究

利用鞘氨醇单胞菌对丁苯橡胶(SBR)胶粉进行脱硫,对比研究SBR胶粉和脱硫SBR胶粉填充SBR胶料的性能。结果表明:鞘氨醇单胞菌不仅可使硫交联键断裂,也可使橡胶主链上的部分碳-碳双键断裂。与SBR胶粉填充的SBR胶料相比,脱硫SBR胶粉填充SBR胶料的转矩和表观交联密度减小,拉伸强度和拉断伸长率增大,邵尔A型硬度和撕裂强度保持不变,阿克隆磨耗量减小,脱硫SBR胶粉与SBR基体结合较紧密,动态滞后损失较小。     

肉桂内生菌Sphingomonas sp.Z45生物转化肉桂醇生成天然2-苯乙醇

从新鲜肉桂枝中筛选出一株高效转化肉桂醇合成天然2-苯乙醇的内生菌,经鉴定为鞘氨醇单胞菌,并命名为Sphingomonas sp. Z45。采用GC-MS法对代谢中间体进行跟踪监测,推测了其可能的代谢途径为:肉桂醇先被氧化、脱羧生成苯乙醛,苯乙醛加氢还原得到产物2-苯乙醇。考察了单因素实验对该生物转化体系的影响,得到反应优化工艺条件为:初始pH=7,接种量5%,三角瓶(150 mL)中装液量为20 mL,菌体培养24 h后,加入底物肉桂醇质量浓度2.5 g/L,在30℃、转速为200 r/min的摇床反应12 h。在该优化条件下,肉桂醇转化率为59.16%,2-苯乙醇质量浓度达到1.48 g/L。     

海藻酸钙包埋枝孢霉对水中Cu2+吸附性能研究

利用海藻酸钙包埋枝孢霉对水中Cu2+吸附性能进行研究。实验结果表明:当海藻酸钙质量分数为3%,CaC l2质量分数为4%,菌体积分数为15%时,包埋制得的固定化小球具有较好的机械性能和较高的吸附量,生物吸附平衡时间3 h。固定化空白小球和活菌的最佳pH值分别为3.5和4.0。在质量浓度为30—500 mg/L时,吸附过程较好地符合Langmu ir吸附模型。在浓度为0.1 mol/L的多种解吸剂中,HNO3解吸效果最好,解吸率达到93.84%。包埋小球重复利用3次,吸附性能没有明显变化。     

Optimization of chitosanase production from Bacillus sp. RKY3 using statistical experimental designs

BACKGROUND: The culture medium and fermentation conditions for the production of constitutive chitosanase from a newly isolated Bacillus sp. RKY3 were optimized statistically. RESULTS: The variables significantly influencing both chitosanase production and cell growth were screened through the Plackett–Burman design, by which maltose, beef extract, MgSO₄, and incubation time were identified as the most significant variables. The optimum values of the selected variables and their mutual interactions were determined through the steepest ascent method and Box–Behnken experimental design. The results demonstrated that 62.30 U mL^?1 chitosanase activity was predicted with optimum conditions of maltose (30.18 g L^?1), beef extract (15.25 g L^?1), MgSO₄ (0.26 g L^?1), and incubation time (50.02 h). The predicted response was verified by the validation experiments, and the optimum conditions resulted in a maximum chitosanase activity of 63.53 ± 1.22 U mL^?1. CONCLUSION: The optimization of fermentation variables resulted in an approximately 11.3‐fold increase in chitosanase activity compared with that observed under unoptimized conditions (from 5.63 U mL^?1 to 63.53 U mL^?1). Copyright © 2009 Society of Chemical Industry     

Sequence and homology model of 3-isopropylmalate dehydrogenase from the psychrotrophic bacterium Vibrio sp. I5 suggest reasons for thermal instability

The leuB gene from the psychrotrophic strain Vibrio sp. I5 has been cloned and sequenced. The gene codes for 3-isopropylmalate dehydrogenase, a 360-residue, dimeric enzyme involved in the biosynthesis of leucine. Three recently solved homologous isopropylmalate dehydrogenase (IPMDH) crystal structures from thermophilic and mesophilic organisms have been used to build a homology model for the psychrotrophic IPMDH and to deduce the possible structural reasons for its decreased thermostability. According to our model the psychrotrophic IPMDH contains fewer stabilizing interactions than its mesophilic and thermophilic counterparts. Elements that have been identified as destabilizing in the comparison of the psychrotrophic, mesophilic and thermophilic IPMDHs are a smaller number of salt-bridges, a reduction in aromatic-aromatic interactions, fewer proline residues and longer surface loops. In addition, there are a number of substitutions of otherwise strictly conserved residues that can be linked to thermostability.      

Production of eicosapentaenoic acid by a recombinant marine cyanobacterium, Synechococcus sp.

The eicosapentaenoic acid (EPA) synthesis gene cluster from an EPA-producing bacterium, Shewanella sp. SCRC-2738, was cloned into a broad-host range vector, pJRD215, and then introduced into a marine cyanobacterium, Synechococcus sp. NKBG15041c, by conjugation. The transconjugant cyanobacteria produced 3.7±0.2% (2.24±0.13 mg/L) EPA (n-3) and 2.5 ±0.2% (1.49±0.06 mg/L) eicosatetraenoic acid (n-3) of the total fatty acids when the cells were cultured at 23°C at a light intensity of 1,000–1,500 Lux. The EPA and eico-satetraenoic acid contents of the cells were increased to 4.6±0.6% (3.86±1.11 mg/L) and 4.7±0.3% (3.86±0.82 mg/L), and 7.5±0.3% (1.76±0.10 mg/L) and 5.1±0.2% (1.19±0.06 mg/L) when they were cultured at low temperature (18°C) and at lower light intensity (40 Lux), respectively.     

Biodegradation of phenol at high concentration by a novel bacterium: Gulosibacter sp. YZ4

BACKGROUND: A novel bacterial strain, Gulosibacter sp. YZ4, has been isolated from activated sludge. Its application potential for phenol biodegradation has not yet been reported, therefore, in this study, biodegradation tests using strain YZ4 were executed under different conditions. RESULTS: The strain was identified as a new member of the genus Gulosibacter and nominated as Gulosibacter sp. YZ4. Phenol biodegradation tests showed that strain YZ4 could thoroughly biodegrade 1000 mg L^?1 phenol across a wide temperature range from 10 to 42 °C and pH range 5 to 11. Degradation of 1000 mg L^?1 phenol was not inhibited by the coexistence of p‐cresol or quinoline. During phenol degradation, strain YZ4 excreted both phenol hydroxylase and catechol 1,2‐dioxygenase to efficiently metabolize phenol. At 36 °C, pH 7.5, strain YZ4 could effectively degrade phenol at concentrations as high as 2000 mg L^?1 within 76 h. Haldane's model with the parameters obtained from the experiments could successfully describe the behavior of the phenol biodegradation by the strain YZ4. CONCLUSIONS: The strain YZ4 has a high potential for applications in phenol wastewater treatment in view of its adaptability to temperature and pH fluctuations and great tolerance to other coexistent toxics. Copyright © 2011 Society of Chemical Industry     

Metal salts requisite for the production of eicosapentaenoic acid by a marine bacterium isolated from mackerel intestines

Metal salts important for the growth and 5,8,11,14,17- ciseicosapentaenoic acid EPA) production of a bacterium isolated from Pacific mackerel intestines were investigated at 25°C in culture media containing 1.0% peptone and 0.50% yeast extract, and the composition of an optimum culture medium was determined. This bacterium could grow in the media in which sodium chloride was the sole added inorganic component. By raising the concentration of sodium chloride from 1.2 to 2.4%, the yield of bacterial cells increased and the yield of EPA reached a maximum at 2.0% NaCl concentration. In contrast to calcium chloride, potassium chloride and magnesium chloride as second metal salts promoted the growth of this bacterium at relatively low concentrations without inhibiting the accumulation of EPA. The yield of EPA reached its maximum value of 51.9 mg/L of culture broth at 8 hr at 2.0% NaCl, 0.15% KCl and 0.16% MgCl₂ concentrations. This yield of EPA was 20% greater than that obtained with Jamarin S artificial sea water. *To whom correspondence should be addressed at Department of Chemical Engineering, Tokyo Institute of Technology, 2-12-1 Ookayama, Meguro-ku, Tokyo 152, Japan. ¹Sagami Chemical Research Center, 4-4-1 Nishi-ohnuma, Sagamihara, Kanagawa 229, Japan.     

一株聚乙烯醇降解菌的降解特性

从腐败的PVA胶水中分离到一株能高效降解并矿化聚乙烯醇的细菌Xa-5,经初步鉴定属于黄单胞菌属（Xanthomonas sp.）.该菌株生长需要复杂的有机氮,蛋白胨和牛肉膏可提高该菌对聚乙烯醇的降解速率,其中蛋白胨的加量对降解聚乙烯醇影响较大.该菌降解聚乙烯醇要求的pH范围较宽.不同聚合度的聚乙烯醇对其降解速率影响不大.该菌的降解过程可能是聚乙烯醇首先吸附在细胞表面,然后进行氧化降解.在静息细胞降解过程中,以TOC表示,1 g•L^-1（DCW）菌体可在52 h内将1 g•L^-1聚乙烯醇基本矿化,但该菌株的PVA氧化酶酶活、脱氢酶酶活和PVA降解总酶活则较难测出.     

Production and stability studies of the biosurfactant isolated from marine Nocardiopsis sp. B4

A potential biosurfactant producing strain, marine Nocardiopsis B4 was isolated from the West coast of India. Culture conditions involving variations in carbon and nitrogen sources were examined at constant pH, temperature and revolutions per min (rpm), with the aim of increasing productivity in the process. The biosurfactant production was followed by measuring the surface tension, emulsification assay and emulsifying index E24. Enhanced biosurfactant production was carried out using olive oil as the carbon source and phenyl alanine as the nitrogen source. The maximum production of the biosurfactant by Nocardiopsis occurred at a C/N ratio of 2:1 and the optimized bioprocess condition was pH 7.0, temperature 30° C and salt concentration 3%. The production of the biosurfactant was growth dependent. The surface tension was reduced up to 29 mN/m as well as the emulsification index E24 was 80% in 6 to 9 days. Properties of the biosurfactant that was separated by acid precipitation were investigated. The biosurfactant activity was stable at high temperature, a wide range of pH and salt concentrations thus, indicating its application in bioremediation, food, pharmaceutical and cosmetics industries.     

Biosorption of mercury and lead by aqueous Streptomyces VITSVK9 sp. isolated from marine sediments from the bay of Bengal,India

Toxic heavy metals are increasingly accumulating in the environment worldwide and are considered to be life threatening contaminants. The biosorption of mercury and lead by marine actinomycetes isolated from marine sediment collected from the Bay of Bengal coast of Puducherry, India, was evaluated. The maximum tolerance concentration (MTC) of Streptomyces sp. was determined by a well diffusion method and a broth dilution method. The effects of the initial metal ion concentration, the pH and the biomass dosage on the biosorption of mercury and lead ions were investigated. The MTC of the isolate to metals was 200 mg·L^-1 for mercury and 1800 mg·L^-1 for lead. At neutral pH, the isolate had a maximum biosorption of metal ions of 200 mg·L^-1 and 150 mg·L^-1 for mercury and lead respectively. Fourier transform infrared (FTIR) absorption spectra showed the chemical interactions between the functional groups in the biomass such as hydroxyl (-OH), amine (-NH₂), carboxyl (-COOH) and the metal ions. The isolate was further characterized by molecular taxonomy and identified as a member of the genus Streptomyces. Based on the phenotypic and phylogenetic analysis, the strain was classified as a new species of the genus Streptomyces and designated as Streptomyces VITSVK9 sp. (HM137310). A blast search of the 16S rDNA sequence of the strain showed the most similarity (95%) with Streptomyces sp. A515 Ydz-FQ (EU384279). Based on the results, it can be concluded that this marine Streptomyces could be used as a biosorbent for the removal of heavy metal ions from aqueous environments.     

Production of lipase from Penicillium sp. using waste oils and Nopalea cochenillifera

The present communication deals with the production of lipase from Penicillium sp. using waste oils and palm cactus (Nopalea cochenillifera) especially as nutrient source of low cost. Two different waste oils were tested: waste frying oil from an industrial kitchen and waste lubricating oil (WLO) from a gas station. Using Doehlert experimental design and response surface methodology, the optimum conditions for lipase production were 96?h fermentation, WLO as the inductor, with specific activity of 0.22?UA?mg^?1. The enzyme was able to remain with more than 58% of its original activity until 30?min at 60°C. The kinetic constants were K_m?=?9.93?mM and V_max?=?2.58 UA min^?1 using p-nitrophenyl palmitate (p-NPP) as substrate. Results showed that Penicillium sp. was able to produce lipase from waste oils using N. cochenillifera, thus having biotechnological potential in waste oil biotransformation.     

Uptake of Cr (VI) by Sphingomonas paucimobilis Biomass from Aqueous Solutions

The Sphingomonas paucimobilis biomass has been successfully utilized to degrade several persistent organic pollutants (POPs). However, few studies have been conducted to use it to remove heavy metals from aqueous solutions. In the present study, biosorption experiments for Cr (VI) were investigated using nonliving biomass of S. paucimobilis isolated from activated sludge, Lianyungang Dapu sewage treatment plant, China. The effects of several parameters including solution pH, contact time, and ionic strength, etc. on Cr (VI) uptake were studied. The biomass was characterized by scanning electron microscopy and energy dispersive X-ray spectroscopy (SEM-EDS) and Fourier transform infrared spectrometer (FTIR). The applicability of the Langmuir and Freundlich models was tested. The correlation coefficients (R) of both models were greater than 0.95. The maximum adsorption capacities were found to be 28.5 mg/g for Cr (VI) at 20°C. The adsorption process was quick and found to follow the pseudo-second-order equation. The optimum adsorption was achieved at pH 2. The adsorption was also NaCl concentrations dependent.