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.     

Characterization of lipoxygenase extracts from Penicillum sp.

Biomasses of Penicillium camemberti and P. roqueforti strains were grown and harvested after 10 d of incubation, a period that corresponded to the maximal dry weight of mycelium as well as to lipoxygenase (LOX) activity. Partially purified LOX extracts were obtained by ammonium sulfate precipitation of the crude enzymatic extracts that had been recovered from the biomasses. The partially purified LOX extracts exhibited a preferential specificity toward free fatty acids, including linoleic, linolenic, and arachidonic acids, rather than fatty acid acylglycerols, including mono-, di-, and trilinolein. The K _m and V _max values of LOX activity were investigated. Normal-phase high-performance liquid chromatography (HPLC) and gas chromatography/mass spectrometry analyses showed that the LOX activity of the microbial extracts converted linoleic acid mainly into the corresponding 9- and 13-hydroperoxides (HPOD). However, the production of a significant proportion of 10-HPOD, ranging from 4 to 9% of the total HPOD, was also demonstrated. In addition, the enantiomeric ratios of the 9- and 13-HPOD produced were determined at both pH optima by chiral-phase HPLC. The results indicated that an almost racemic mixture had been obtained which can be related to either low enantioselectivity of LOX or the presence of isozymes showing complementary enantioselectivity.     

Biodegradation ofVernonia galamensis seed Oil byAcinetobacter andPseudomonas sp.

The biodegradability ofVernonia galamensis seed oil (VO) has been demonstrated with two environmental bacterial strains,Acinetobacter Iwoffi (HU 3955), andPseudomonas sp. (HU 4020). A time-dependent increase in the degradative activities of both bacteria species was apparent. There wasca. 60% decrease in the amount of VO over an eightday incubation period. Additionally, lipolytic activity was evident from the amount of free fatty acid (FFA) that was generated. The percent FFA of the residual oil were 82% for thePseudomonas strain, and 62% for theAcinetobacter strain. The weight per epoxy value of the VO in the fermentation medium remained relatively constant over the incubation period, suggesting the lack of preference for either the epoxidized or nonepoxidized acids present in VO.     

Isolation of a new highly CO2 tolerant fresh water MicroalgaChlorella sp. KR-1

A fresh water microalga, which has tolerance to high concentrations of CO₂, was isolated. The KR-1 strain was identified as a genusChlorella. ThoughChlorella KR-1 showed maximum growth at 10 % (v/v) CO₂, the strain showed a good growth rate up to 50 % (v/v) CO₂. The results indicated the feasibility of the KR-1 strain for massive cultivation using condensed stack gases.     

Pseudomonas sp. lipase-catalyzed synthesis of geranyl esters by transesterification

Pseudomonas sp. lipase was immobilized by adsorption onto five supports and tested for its ability to synthesize geranyl esters by transesterification using short-chain triacylglycerols as acyl donors. Reaction mixtures were prepared in 2 mL ofn-hexane, 0.1 M geraniol, 0.03M triacylglycerol, and 200 units of lipase, and incubated at 30°C and 200 rpm for 24 h. Overall, glass beads were the best support. Geranyl acetate and caproate performed best with Duolite (77.5 and 95.3%, respectively). Geranyl butyrate and caprylate performed best with polyvinylpyrrolidone, (80.2 and 95.5%, respectively). Values for nonimmobilized enzyme also were obtained. Immobilization improved yields, with geranyl caproate exhibiting best results.     

Production of eicosapentaenoic acid bySaprolegnia sp. 28YTF-1

Saprolegnia sp. 28YTF-1, isolated from a freshwater sample, is a potent producer of 5,8,11,14,17-cis-eicosapentaenoic acid (EPA). The fungus used various kinds of carbon sources, such as starch, dextrin, sucrose, glucose, and olive oil for growth, and olive oil was the best carbon source for EPA production. The EPA content reached 17 mg/g dry mycelium (0.25 mg/L) when the fungus was grown in a medium that contained 2.5% olive oil and 0.5% yeast extract, at pH 6.0 and 28°C for 6 d with shaking. Accompanying production of arachidonic acid (AA; 3.2 mg/g dry mycelia, EPA/AA = 5.1) and other ω6 polyunsaturated fatty acids was low. Both EPA content and EPA/AA ratio increased in parallel by lowering growth temperature. Triglyceride was the major mycelial lipid (ca. 84%), but EPA comprised only 2.2% of the total fatty acids of this lipid. About 40% of the EPA produced was found in polar lipids, such as phosphatidylethanolamine (EPA content, 28.2%), phosphatidylcholine (13.6%), and phosphatidylserine (21.2%).     

Studies on Bioflocculant Production by Arthrobacter sp. Raats, a Freshwater Bacteria Isolated from Tyume River, South Africa

A bioflocculant-producing bacteria was isolated from Tyume River in the Eastern Cape Province, South Africa and identified by 16S rRNA gene nucleotide sequence to have 91% similarity to Arthrobacter sp. 5J12A, and the nucleotide sequence was deposited in GenBank as Arthrobacter sp. Raats (accession number HQ875723). The bacteria produced an extracellular bioflocculant when grown aerobically in a production medium containing glucose as sole carbon source and had an initial pH of 7.0. Influences of carbon, nitrogen and metal ions sources, as well as initial pH on flocculating activity were investigated. The bacteria optimally produced the bioflocullant when lactose and urea were used as sole sources of carbon and nitrogen respectively with flocculating activities of 75.4% and 83.4% respectively. Also, the bacteria produced the bioflocculant optimally when initial pH of the medium was 7.0 (flocculating activity 84%), and when Mg(2+) was used as cation (flocculating activity 77%). Composition analyses indicated the bioflocculant to be principally a glycoprotein made up of about 56% protein and 25% total carbohydrate.     

蓝细菌鱼腥藻7120混合营养培养的生长和生理特征

Mixotrophic growth is one potential mode for mass culture of microalgae and cyanobacteria particularly suitable for the production of high value bioactive compounds and fine chemicals.The typical heterocystous cyanobacterium Anabaena sp.PCC 7120 was grown in the presence of exogenous glucose in light.Glucose improved the cell growth evidently,the maximal specific growth rate under mixotrophic condition(0.38 d1)being 1.6-fold of that of photoautotrophic growth.Mixotrophy caused a variation in cellular pigment composition,increasing the content of chlorophyll a and decreasing the contents of carotenoid and phycobiliprotein relative to chlorophyll a.Fluorescence emission from photosystem II(PSII)relative to photosystem I was enhanced in mixotrophic cells,implying an increased energy distribution in PSII.Glucokinase(EC 2.7.1.2)activity was further induced in the presence of glucose.The mixotrophic culture was scaled up in a 15 L airlift photobioreactor equipped with an inner and an outer light source.A modified Monod model incorporating the specific growth rate and the average light intensity in the reactor was developed to describe cell growth appropriately.The understanding of mixotrophic growth and relevant physiological features of Anabaena sp.PCC 7120 would be meaningful for cultivation and exploitation of this important cyanobacterial strain.     

Rhizobium sp.N613合成胞外多糖的发酵动力学研究

对Rhizobium sp.N613合成胞外多糖(REPS)的分批发酵过程和发酵动力学进行了研究.应用Logistic方程和Luedeking-Piret方程,得到了描述Rhizobium sp.N613菌体生长、REPS合成和底物消耗的动力学模型.模型反映了该菌株合成REPS分批发酵过程的动力学特征,模型值与实验数据拟合良好.对Rhizobium sp.N613发酵动力学模型和合成REPS的生理学特性进行了分析,优化了合成REPS的发酵工艺,补料分批发酵实验结果表明,REPS产量从8.21 g·L-1提高到 11.31 g·L-1.     

Purification and characterization of a thermophilic chitinase produced by <Emphasis Type="Italic">Aeromonas</Emphasis> sp. DYU-Too7

An extracellular chitinase, produced by Aeromonas sp. DYU-Too7, was purified in the following procedures: ammonium sulfate precipitation, ultrafiltration, chromatographic separation of DEAE-sepharose CL-6B and sephacrylS-100HR. The resulting chitinase has a molecular mass of 36 kDa, an optimal reaction pH of 5.0, and an optimal reaction temperature of 70°C. It retains almost 100% activity in the pH range of 5.0–8.0. This chitinase has a high thermal tolerance and retained 90% of its activity at 50°C and 75% at 60°C. Enzyme activity was inhibited by Ba²⁺, Hg²⁺, Mg²⁺ and Ag⁺ cations, but was not substantially inhibited by the K⁺ cation nor the chelating agent EDTA. The K _m and V _mα , using colloidal chitin as a substrate, are 6.3 g/L and 18.69 μmol/min/mg-protein, respectively. The 36 kDa chitinase of Aeromonas sp. DYU-Too7 is an exo-type enzyme, because chitobiose was the main hydrolysate in hydrolysis of colloidal chitin.     

A physiological study on growth and dibenzothiophene (DBT) desulfurization characteristics of Gordonia sp. CYKS1

Physiological characteristics of DBT desulfurization and cell growth ofGordonia sp. CYKS1 were investigated. It exhibited a preference to ethanol in a medium containing two carbon sources, ethanol and one of the carbohydrates used, glucose, sucrose, maltose, and galactose although it consumed both carbon sources simultaneously. Cell growth on ethanol or glucose followed the Monod kinetics. The optimal range of pH for the desulfurization of DBT and the cell growth was 7 to 8. The desulfurization rate decreased about 30% at pH 6, and no significant desulfurization or cell growth was observed at pH 5. As the initial DBT concentration increased up to 1.5 mM, the desulfurization rate also increased while no significant changes in the growth rate were observed. The maximum desulfurization rate was 12.50 μmol L^-1 h^-1 at an initial DBT concentration of 1.5 mM. Cell growth and desulfurization activity were severely inhibited by the presence of 2-hydroxybiphenyl (2-HBP). When 0.05 mM of 2-HBP was added at the beginning, both of the desulfurization rate and cell growth rate decreased about 20%. It was found that cell growth and desulfurization were completely inhibited in the presence of 2-HBP at 0.15 mM or a higher concentration. The inhibition by 2,2′-dihydroxybiphenyl (DHBP) was less severe than 2-HBP. About 80% of desulfurization activity was retained in the presence of 2,2′-DHBP at 0.4 mM.     

Alarm response of the marine mud snail,Nassarius obsoletus: Specificity and behavioral priority

Nassarius obsoletus, N. vibex, andN. trivittatus were exposed to juices of conspecifics and taxonomically and ecologically related gastropod species. Conspecific juices caused the strongest alarm responses, while those from taxonomically related species (congeners) induced less pronounced responses. Juices from nonrelated species induced no alarm response with one notable exception, the strong alarm response ofN. trivittatus toUrosalpinx cinerea juices. Nonrelated gastropod species induced stronger feeding responses than more closely related species. The strongest feeding responses, however, were induced by juices from nongastropod mollusks. The apparent balance between feeding attraction to less related species and alarm response to more related species was further examined by manipulating the snails' diet. Starvation resulted in a great reduction of conspecific alarm responsiveness. Following the resumption of feeding, the original responsiveness was again observed. Although the alarm response is likely an antipredator device, conclusive evidence for its effectiveness in reducing predation is lacking. One of the predators ofNassarius obsoletus which may be responsible for maintenance of an alarm response is the green crab,Carcinus maenas.This paper is based in part upon a thesis submitted by D.S. to Boston University for the degree of Master of Arts.     

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     

鱼腥藻 Anabaena sp. PCC7120的混合营养生长

在高光强为１６０μＥ／（ｍ２·ｓ）、低光强为１６μＥ／（ｍ２·ｓ）、葡萄糖浓度０～３０ｇ／Ｌ范围内,、进行了鱼腥藻Ａｎａｂａｅｎａｓｐ．ＰＣＣ７１２０的摇瓶光自养和混合营养培养．在高光强下最大藻细胞密度（０．９２～３．１ｇ／Ｌ）明显高于低光强（０．１１～０．５８ｇ／Ｌ）,而且高光强使混合营养培养的对数期缩短．在不同光强下,葡萄糖浓度在０～１８ｇ／Ｌ范围内提高显著促进了细胞的生长,在１８～３０ｇ／Ｌ范围内变化对细胞生长不再有更大的影响．高光强促进了藻细胞对葡萄糖的利用．在高光强下随着葡萄糖浓度的提高,细胞得率逐渐变小．     

Chemical Defenses of the Sacoglossan Mollusk Elysia rufescens and Its Host Alga Bryopsis sp.

Sacoglossans are a group of opisthobranch mollusks that have been the source of numerous secondary metabolites; however, there are few examples where a defensive ecological role for these compounds has been demonstrated experimentally. We investigated the deterrent properties of the sacoglossan Elysia rufescens and its food alga Bryopsis sp. against natural fish predators. Bryopsis sp. produces kahalalide F, a major depsipeptide that is accumulated by the sacoglossan and that shows in vitro cytotoxicity against several cancer cell lines. Our data show that both Bryopsis sp. and Elysia rufescens are chemically protected against fish predators, as indicated by the deterrent properties of their extracts at naturally occurring concentrations. Following bioassay-guided fractionation, we observed that the antipredatory compounds of Bryopsis sp. were present in the butanol and chloroform fractions, both containing the depsipeptide kahalalide F. Antipredatory compounds of Elysia rufescens were exclusively present in the dichloromethane fraction. Further bioassay-guided fractionation led to the isolation of kahalalide F as the only compound responsible for the deterrent properties of the sacoglossan. Our data show that kahalalide F protects both Bryopsis sp. and Elysia rufescens from fish predation. This is the first report of a diet-derived depsipeptide used as a chemical defense in a sacoglossan.     

Fatty Acid Desaturation and Elongation Reactions of <Emphasis Type="Italic">Trichoderma</Emphasis> sp. 1-OH-2-3

The fatty acid desaturation and elongation reactions catalyzed by Trichoderma sp. 1-OH-2-3 were investigated. This strain converted palmitic acid (16:0) mainly to stearic acid (18:0), and further to oleic acid (c9-18:1), linoleic acid (c9,c12-18:2), and α-linolenic acid (c9,c12,c15-18:3) through elongation, and Δ9, Δ12, and Δ15 desaturation reactions, respectively. Palmitoleic acid (c9-16:1) and cis-9,cis-12-hexadecadienoic acid were also produced from 16:0 by the strain. This strain converted n-tridecanoic acid (13:0) to cis-9-heptadecenoic acid and further to cis-9,cis-12-heptadecadienoic acid through elongation, and Δ9 and Δ12 desaturation reactions, respectively. trans-Vaccenic acid (t11-18:1) and trans-12-octadecenoic acid (t12-18:1) were desaturated by the strain through Δ9 desaturation. The products derived from t11-18:1 were identified as the conjugated linoleic acids (CLAs) of cis-9,trans-11-octadecadienoic acid and trans-9,trans-11-octadecadienoic acid. The product derived from t12-18:1 was identified as cis-9,trans-12-octadecadienoic acid. cis-6,cis-9-Octadecadienoic acid was desaturated to cis-6,cis-9,cis-12-octadecatrienoic acid by this strain through Δ12 desaturation. The broad substrate specificity of the elongation, and Δ9 and Δ12 desaturation reactions of the strain is useful for fatty acid biotransformation.     

Biosynthetic pathway of diepoxy bicyclic FA from linoleic acid by Clavibacter sp. ALA2

The biosynthetic pathway of two bicyclic FA, 12∶17, 13∶17-diepoxy-9(Z)-octadecenoic acid (DEOA) and 7-hydroxy-12∶17, 13∶17-diepoxy-9(Z)-octadecenoic acid (hDEOA), by Clavibacter sp. ALA2 was investigated. When cultivated with linoleic acid as a substrate, the strain produced 12,13,17-trihydroxy-9(Z)-octadecenoic acid (THOA), DEOA, and hDEOA as well as other FA. To clarify the synthetic route to these bicyclic FA, the strain was cultivated with purified THOA as a starting substrate. THOA was consumed almost completely by the strain with sequential generation of DEOA and hDEOA. Moreover, the strain produced hDEOA when cultivated with purified DEOA. Therefore, it was confirmed that THOA was a precursor of these bicyclic FA and that hDEOA was generated from DEOA. Based on our previously reported result that linoleic acid is first converted to 12,13-dihydroxy-9(Z)-octadecenoic acid (DHOA) and the present results, the overall biosynthetic pathway for the diepoxy bicyclic FA from linoleic acid was postulated as: linoleic acid→DHOA→THOA→DEOA→hDEOA.     

A new cucurbitacin profile forCucurbita andreana: A candidate for cucurbitacin tissue culture

In addition to previously reported cucurbitacins B,1, and D,2, cucurbitacin E,3, and I,4, aglycones and their glucosides 2-O--glucopyranosyl-cucurbitacin E,5, and 2-O--glucopyranosyl-cucurbitacin I,6, were isolated and identified as constituents ofCucurbita andreana on the basis of MS, FD-MS, 1 H NMR and¹³C NMR spectroscopy. Also, 2-O--glucopy-ranosyl-cucurbitacin B, 7, cucurbitacin B glucoside was isolated and identified.     

新筛选菌种Sphingopyxis sp.USTB-05对微囊藻毒素的生物降解

A promising bacterial strain for the biodegradation of Microcystins (MCs) was isolated from Dianchi lake in China and identified as Sphingopyxis sp. USTB-05 by the analysis of 16s rDNA. Initial MC-RR of 42.3 mg&;#8226;L－1 was completely degraded by USTB-05 within 36 h, which was a relatively high biodegradation rate of MC-RR. With the cell-free extract (CE) of Sphingopyxis sp. USTB-05, MC-RR was biodegraded at a more rapid biodegradation rate compared with its strain, so that initial MC-RR of 42.3 mg&;#8226;L－1 was completely biodegraded within 10 h. During the bio-reaction of MC-RR catalyzed by CE, two intermediate metabolites and a dead-end product of MC-RR were observed on HPLC profiles and all of them had similar scanning profiles in the wavelength from 200 to 300 nm, indicating that the group of Adda in all products of MC-RR remained intact.     

Sorption of boron by invasive marine seaweed: Caulerpa racemosa var. cylindracea

The sorption of boron from aqueous solution onto Caulerpa racemosa var. cylindracea (CRC), collected from Seferihisar/Izmir region in Turkey, was investigated as a function of pH, temperature, initial boron concentration, adsorbent dosage, contact time and ionic strength. Optimum conditions for the sorption of boron were obtained at pH 7.5, 318 K, 8 mg L⁻¹ initial boron concentration, 0.2 g of CRC, 2.5 h contact time and greater ionic strength (10⁻¹ M NaCl). As the temperature was increased the boron removal took place with higher percentages. In experiments conducted at optimum conditions, maximum boron sorption was determined to be about 63%. The experimental data were analyzed by Freundlich, Langmuir and Dubinin–Radusckevich (DR) equations. Freundlich and DR models provide best conformity with the experimental data. In order to describe kinetics of boron sorption onto CRC, first-order Lagergren equation, pseudo-second-order kinetic model and intraparticle diffusion model were used. It was seen that the first order Lagergren equation was better described than the pseudo-second-order kinetic model. Thermodynamic parameters of sorption process were also calculated. It was obtained that sorption process was not spontaneous. The characterization of CRC was carried out by Fourier transform infrared spectroscopy (FTIR) analysis.