Production and characterization of glycolipid biosurfactant from Streptomyces enissocaesilis HRB1 and its evaluation for biomedical and bioremediation applications

The aim of this investigation is to produce and characterize biosurfactant from Streptomyces sp. HRB1 and to evaluate its biomedical and bioremediation potential. Biosurfactant producing property of Streptomyces sp. HRB1 isolated from petroleum contaminated soil was confirmed by hemolytic and oil spread assays. Based on the results of FT-IR spectral and GC–MS analysis, the biosurfactant was confirmed as glycolipid type. Biosurfactant from Streptomyces sp. HRB1 exhibited 71% inhibition against Pseudomonas aeruginosa biofilm formation, 77.33% quorum sensing inhibition property against Chromobacterium violeceum MTCC 2656, more than 80% inhibition in antioxidant assays namely, DPPH, ABTS, and metal chelation, promising anti-proliferative activity against leukemia and myeloma cells with low IC₅₀ values, 96% decolorization of malachite green within 48 h of reaction time, and minimal toxicity against normal cell lines in dose-dependent manner. The taxonomic position of the potential strain HRB1 was further confirmed as Streptomyces enissocaesilis HRB1 based on their phenotypic and molecular characteristics. To conclude, Streptomyces enissocaesilis HRB1 isolated from petroleum-contaminated soil is a promising source for low-cost production of glycolipid biosurfactant with potential biomedical and environmental applications such as antiphytofungal, antibiofilm, anti-quorum sensing, antioxidant, anticancer, and dye degradation properties.     

氮源对利迪链菌素生产及相关次级代谢物分布的影响

The effects of nitrogen sources on streptolydigin production and distribution of secondary metabolites were investigated for flask cultured S.lydicus AS 4.2501.When peptone,asparamide,and glutamic acid were ex- amined as the nitrogen source,respectively,liquid chromatography-mass spectrometry（LC-MS）and photodiode array（PDA）analyses revealed the formation of two analogues of streptolydigin in the fermentation broth.When soybean meal was used as the source of nitrogen,three analogues of streptolydigin were detected.The use of am- monium sulfate as a source of nitrogen resulted in a lower pH value of the fermentation system,thus inhibiting streptolydigin biosynthesis and changing the metabolic profiling.Among the nitrogen sources that were made use of,glutamic acid was most favorable to the formation of streptolydigin.Simultaneously,this study also showed that the changing nitrogen sources resulted in altering the production and relative ratios of streptolydigin and its analogues.     

Structural Analysis of Cytochrome P450 105N1 Involved in the Biosynthesis of the Zincophore, Coelibactin

Coelibactin is a putative non-ribosomally synthesized peptide with predicted zincophore activity and which has been implicated in antibiotic regulation in Streptomyces coelicolor A3(2). The coelibactin biosynthetic pathway contains a stereo- and regio-specific monooxygenation step catalyzed by a cytochrome P450 enzyme (CYP105N1). We have determined the X-ray crystal structure of CYP105N1 at 2.9 Å and analyzed it in the context of the bacterial CYP105 family as a whole. The crystal structure reveals a channel between the α-helical domain and the β-sheet domain exposing the heme pocket and the long helix I to the solvent. This wide-open conformation of CYP105N1 may be related to the bulky substrate coelibactin. The ligand-free CYP105N1 structure has enough room in the substrate access channel to allow the coelibactin to enter into the active site. Analysis of typical siderophore ligands suggests that CYP105N1 may produce derivatives of coelibactin, which would then be able to chelate the zinc divalent cation.     

The butenolide signaling molecules SRB1 and SRB2 induce lankacidin and lankamycin production in Streptomyces rochei

New signaling molecules that induce lankacidin and lankamycin production in Streptomyces rochei were extracted from the culture filtrate and purified by Sephadex LH20 and silica gel chromatography with the help of bioassay. Chiral HPLC and ESI-MS analyses indicated the presence of two active components--SRB1 and SRB2--and their molecular formulas were established to be C15H24O5 and C16H26O5, respectively. By extensive NMR analysis, SRB1 and SRB2 were determined to be 2-(1'-hydroxy-6'-oxo-8'-methylnonyl)-3-methyl-4-hydroxybut-2-en-1,4-olide and 2-(1'-hydroxy-6'-oxo-8'-methyldecyl)-3-methyl-4-hydroxybut-2-en-1,4-olide, respectively. These structures were finally confirmed by chemical synthesis and the absolute configuration at C-1' was determined to be R in each case. The synthetic 1'R isomers induced production of lankacidin and lankamycin at around 40 nM concentrations. SRB1 and SRB2 are therefore distinct from the well-known 2,3-disubstituted γ-butyrolactone molecules such as A-factor, virginia butanolide, and SCB1 and and belong, like avenolide, recently isolated from Streptomyces avermitilis, to the γ-butenolide family.     

枝链霉菌L2001木聚糖酶的化学修饰及其活性中心

分别用多种化学修饰剂,如NBS、WRK对枝链霉菌L2001木聚糖酶进行化学修饰,结果表明,作用于色氨酸的试剂NBS能使酶活明显降低,而作用于谷氨酸、半胱氨酸、赖氨酸、精氨酸、丝氨酸及组氨酸残基的试剂对酶活力影响明显。为了进一步研究色氨酸对酶活力的影响作用,通过测定修饰酶的假一级常数,得到至少有一个色氨酸残基在枝链霉菌L2001木聚糖酶的活性中心,并通过底物对化学修饰的保护试验得到,色氨酸残基处在酶与底物的结合位点。结合荧光光谱再次确定色氨酸处于该酶的活性部位,是保持酶活性的必需基团。     

含新农抗702的HP-10树脂解吸及再生工艺

为探索含新农抗702的HP-10大孔树脂的解吸及树脂再生的适宜条件,采用静态、动态方式对树脂进行洗脱;采用生物学法(管碟法)对树脂洗脱液中的残留物进行检测。结果表明,静态解吸时乙醇能够使含新农抗702的大孔HP-10树脂一次性解吸率达到90%;动态解吸时洗脱峰比较集中便于后续浓缩;通过比较再生前后树脂吸附的吸附性能发现,树脂基本再生完全,且再生方法简便,可使树脂重新获得吸附性能,故可反复使用,达到节约原料的目的。     

Epsilon-Polylysine Fermentation and its Recovery Using Carboxymethyl Cellulose (CMC)-Conjugated Magnetite

The easily prepared polyanionic magnetic nanoparticle (MNP) was employed to recover a polycationic natural food preservative, epsilon-polylysine (?-PL) from fermentation broth of Streptomyces albulus. ?-PL with a final concentration of 10 g/L could be produced in a 5 day pH controlled two-stage fermentation of S. albulus. The polyanionic MNP was prepared by co-precipitating Fe(II)/Fe(III) solution in the presence of water soluble carboxymethyl cellulose (CMC) at pH 11. The Langmuir isotherm can well describe the adsorption behavior between CMC-conjugated MNP (CMC-MNP) and ?-PL that 0.38 g of ?-PL could be captured by 1 g of CMC-MNP at pH 5, 37°C. Approximately, 80% of ?-PL adsorbed on the freshly prepared CMC-MNP could be eluted by 0.1 N HCl. The adsorption capacity of CMC-MNP remained at a constant level for at least 5 repeated adsorptions. The ?-PL isolated by CMC-MNP not only demonstrated an improved purity but also its antimicrobial activity as compared with α-polylysine against the bacterium E. coli.     

白蚁肠道二株链霉菌株所产纤维素酶的分离、纯化及其特性(英文)

通过硫酸铵沉析、依次过葡聚糖G100和阴离子交换树脂DEAE葡聚糖A50柱,从白蚁(Odontotermesformosanus)肠道2株放线菌株发酵液中获得了1种葡聚糖外切酶(C1)、2种葡聚糖内切酶(CX)和1种β-葡萄糖苷酶(β-glucosidase,βG)。该2株放线菌株在形态、生长、生理生化特性上显示属于链霉菌属种类。SDS-PAGA电泳分析表明上述4种酶的分子质量分别为76.9、22.3、66.2、31.9kD。外切酶在pH 5.6,内切酶和β-葡萄糖苷酶在pH 5.0;以及内切酶和外切酶在50℃,β-葡萄糖苷酶在40℃时,有最大酶活性,甚至温度高达70℃时,这些酶仍有50%以上的酶活性。二价阳离子如Fe2+、Ca2+在质量浓度100mg/L条件下对酶有激活作用,而Mn2+、Cu2+、Zn2+、Co2+对酶活起抑制作用。结果表明来自白蚁肠道2株链霉菌株所产的纤维素酶可能以Fe2+和Ca2+作为辅基,属于酸性、耐热性酶。这类酶具备用于工业上分解不溶性的纤维素的能力。