一株耐盐菌DL41诱导合成Ectoine及海藻糖的研究

从盐池淤泥中采集土样,用盐梯度法筛选分离得到一株耐盐微生物DL41,对其进行菌落及菌体形态、分子生物学鉴定和生长特性研究.通过PCR技术,进行16S rDNA鉴定,建立系统发育树.还研究了耐盐微生物DL41合成相容性物质Ectoine和海藻糖诱导条件.     

一株耐盐菌EC51的生长特性及其系统发育分析

从大连市普兰店盐场盐池底部的淤泥中筛选出一株耐盐菌EC51,研究了分离菌株的形态和生长特性,该菌株为革兰氏阳性杆状菌。以其16S rDNA序列相似性为基础构建系统发育树。结果表明,EC51与Bacillus Pumilus的相似性达到99%。耐盐菌EC51的渗透压补偿溶质(Ec-toine)生成情况实验表明,用含2 mol/L NaCl的肉汤培养基,在30℃条件下培养48 h,诱导生成Ectoine为197.8 mg/L。     

Ectoine对Cu-Zn型SOD酶热稳定性的影响

研究了Ectoine对Cu-Zn型SOD酶在热处理条件下的保护作用.研究结果表明,Ectoine对经热处理的Cu-Zn型SOD酶有明显的保护作用.0.7 mmol/L的Ectoine可使经50、90、100℃处理的Cu-Zn型SOD酶的相对酶活力分别提高25.7%、19.0%、13.2%;0.5 mmol/L的Ectoine在60℃下可使酶活提高26.5%;0.3 mmol/L的Ectoine在80℃下可使酶活提高22.1%,0.1 mmol/L的Ectoine在70℃下可使酶活提高28.6%.Ectoine与脯氨酸、甘油、海藻糖、谷氨酸、甜菜碱,在相同热处理条件下比较,Ectoine对Cu-Zn型SOD酶的热保护作用最为明显,且用量最少.     

Prospective randomized study to evaluate the efficacy and tolerability of Ectoin® containing Eye Spray (EES09) and comparison to the liposomal Eye Spray Tears Again® (TA) in the treatment of dry eye disease

PurposeTo subjectively and objectively evaluate the efficacy and tolerability of preservative-free Ectoin ® Eye Spray – Colloidal (EES09) and Tears Again ® Eye Spray (TA) in subjects with mild-moderate dry eye disease (DED), and to compare efficacy of these two eye sprays with each other.MethodsThirty-six volunteers (average age 32.3 ± 16.1 years; 26 females) were successfully recruited for this prospective double-blind study with between-subject design and randomly divided into two groups (gender and age balanced): Group A received EES09 and Group B received TA during the treatment phase. Inclusioncriteria were a minimum age of 18 years, a score of 18.0 or higher on the OSDI questionnaire, and a non-invasive tear break up time (NIKBUT, Oculus Keratograph M5, Oculus, Germany) of no more than 10s in at least one eye. The objective variables NIKBUT, conjunctival redness, lipid layer and osmolarity (TearLab Cooperation, USA) were assessed at baseline, 10 min. after spray application and after a treatment phase of 10±3 days (3x applications daily). Symptoms, tolerance and handling were evaluated with the OSDI and VAS questionnaires.ResultsA statistically significant increase in NIKBUT and improvement in DED symptoms were obtained for the overall group (mean NIKBUT 7.7 ± 1.7s vs. 11.6 ± 4.6s, p<0.001; mean OSDI score: 36.1 ± 12.7 vs. 20. 7± 12.7, p<0.001) during course of treatment. No statistically significant effect was established for the variables lipid layer (p = 0.406), conjunctival redness (p = 0.766) and osmolarity (p = 0.378). No statistically significant differences were observed between the groups, for any variable. The noninferioritycriterion for EES09 towards TA could be shown for the scores of the dry eye symptoms via VAS questionnaire and the variable NIKBUT.ConclusionsA beneficial treatment effect was confirmed for both, symptoms of DED and the objective variable NIKBUT. Both eye sprays were rated favourably in view of perceived tolerability and handling of the spray bottle.     

一株渗透压补偿溶质Ectoine生成菌的分离及其分子生物学鉴定

从大连普兰店盐场盐池底土壤中筛选获取一株产渗透压补偿溶质Ectoine的耐盐菌株SL39,其16SrDNA序列分析表明,菌株SL39与Halobacillustrueperi具有100%同源性。该菌株最适生长温度为30℃,最适初始pH为7.0,能够在0￣25%(w/v)NaCl浓度范围内生长,最适生长NaCl浓度为5%(w/v)。SL39菌株在2mol/LNaCl的培养基中诱导能合成Ectoine,在30℃诱导培养48h其合成量为304.8mg/L。     

Production and characterization of ectoine by Marinococcus sp. ECT1 isolated from a high-salinity environment

A halophilic bacterium isolated from a salt environment in southern Taiwan was identified as a Marinococcus sp. ECT1. This bacterium could synthesize and accumulate intracellular ectoine as a compatible solute capable of resisting osmotic stress in a hyper-osmotic environment. This study also developed a semi-synthesized medium (YAMS medium), capable of facilitating the growth of this Marinococcus sp. ECT1 with 600 mg/L crude ectoine production. Moreover, Marinococcus sp. ECT1 was grown on YAMS medium containing different initial yeast extract concentrations (C(YE)) (0 to 60 g/L) to demonstrate how C(YE) affects crude ectoine production. While the maximum cell concentration was increased by 23-fold when the C(YE) was 40 g/L, the maximum crude ectoine production reached 2.5 g/L when C(YE) was 40 g/L. In addition to demonstrating the success of the fermentation strategy of ectoine in increasing the production and production yield, experimental results further demonstrated that the fermentation medium of ectoine is highly promising for commercialization. Furthermore, the molecular weight and chemical structure of ectoine were identified and characterized by FAB-MS and (1)H-NMR.     

高产Ectoine耐盐菌株的筛选

对产Ectoine的耐盐菌株进行筛选,在不同盐度条件下进行Ectoine诱导、释放和检测,并对菌种形态进行鉴定。实验过程中,从盐度15%~25%均检测出Ectoine,进行综合比对得出结论 GH07菌株为高产Ectoine耐盐菌株。     

Ectoine对Cu—Zn型SOD酶热稳定性的影响

研究了Ectoine对Cu-Zn型SOD酶在热处理条件下的保护作用．研究结果表明，Ectoine对经热处理的Cu-Zn型SOD酶有明显的保护作用．0．7mmol／L的Ectoine可使经50、90、100℃处理的Cu-Zn型SOD酶的相对酶活力分别提高25．7％、19．0%、13．2％；0．5mmol／L的Ectoine在60℃下可使酶活提高26．5%；0．3mmol／L的Ectoine在80℃下可使酶活提高22．1％，0．1mmol／L的Ectoine在70℃下可使酶活提高28．6％．Ectoine与脯氨酸、甘油、海藻糖、谷氨酸、甜菜碱，在相同热处理条件下比较，Ectoine对Cu-Zn型SOD酶的热保护作用最为明显，且用量最少．     

一株Ectoine生成菌DL031的分离及特性研究

从大连普兰店盐场盐池土壤中分离到一株产渗透压补偿溶质Ectoine的菌株DL031,通过16SrDNA序列分析,菌株DL031与Halomonas marina相似性为99%.菌株DL031可在含有0～200 g/LNaCl培养基中生长,生长最适NaCl浓度为100 g/L.DL031菌株在含有NaCl的培养基中诱导能合成Ectoine;在含3 mol/L NaCl的培养基,30℃诱导培养48 h,Ectoine的合成量为98.8 mg/L.