Phosphatase activity and culture conditions of the yeast Candida mycoderma sp. and analysis of organic phosphorus hydrolysis ability

Orthophosphate is an essential but limiting macronutrient for plant growth. About 67% cropland in China lacks sufficient phosphorus, especially that with red soil. Extensive soil phosphorus reserves exist in the form of organic phosphorus, which is unavailable for root uptake unless hydrolyzed by secretory acid phosphatases. Thus, many microorganisms with the ability to produce phosphatase have been exploited. In this work, the activity of an extracellular acid phosphatase and yeast biomass from Candida mycoderma was measured under different culture conditions, such as pH, temperature, and carbon source. A maximal phosphatase activity of 8.47 × 10^5±0.11× 10^5 U/g was achieved by C. Mycoderma in 36 hr under the optimal conditions. The extracellular acid phosphatase has high activity over a wide pH tolerance range from 2.5 to S.0 （optimum pH 3.5）. The effects of different phosphorus compounds on the acid phosphatase production were also studied. The presence of phytin, lecithin or calcium phosphate reduced the phosphatase activity and biomass yield significantly. In addition, the pH of the culture medium was reduced significantly by lecithin. The efficiency of the strain in releasing orthophosphate from organic phosphorus was studied in red soil （used in planting trees） and rice soil （originating as red soil）. The available phosphorus content was increased by 230% after inoculating 20 days in rice soil and decreased by 50% after inoculating 10 days in red soil. This work indicates that the yeast strain C. mycoderma has potential application for enhancing phosphorus utilization in plants that grow in rice soil.     

Submerged vegetation removal promotes shift of dominant phytoplankton functional groups in a eutrophic lake

Historical data indicate that the dominance of submerged plants in Dianchi Lake in the 1960 s was characterized by low algal density with dominance of non-toxic group J（Scenedesmus,Pediastrum,etc.）. The removal of submerged plants,which began in the 1970 s,resulted in the expansion of bloom-forming Microcystis（group M）. Laboratory experiments suggested that Microcystis aeruginosa was inclined to grow and develop at elevated temperatures. The growth of Scenedesmus obliquus was slower than that of co-cultivated M. aeruginosa in the absence of Ceratophyllum demersum,especially at higher temperatures. The existence of submerged plant C. demersum could inhibit the growth of the harmful algae M. aeruginosa and this inhibitory effect by C. demersum was enhanced with an increase in temperature. Instead,with C. demersum,the growth of S. obliquus was not inhibited,but the co-cultivated M. aeruginosa was eliminated in a short time. Combined with the historical data and laboratory experiments,it was indicated that the submerged plants might play important roles in the dominance of the non-toxic group J in the historical succession. Consequently,the introduction of the submerged plant such as C. demersum might alter the dominant phytoplankton functional groups from M to J and benefit the restoration of the eutrophic lake.     

Community structure and elevational diversity patterns of soil Acidobacteria

Acidobacteria is one of the most dominant and abundant phyla in soil,and was believed to have a wide range of metabolic and genetic functions. Relatively little is known about its community structure and elevational diversity patterns. We selected four elevation gradients from 1000 to 2800 m with typical vegetation types of the northern slope of Shennongjia Mountain in central China. The vegetation types were evergreen broadleaved forest,deciduous broadleaved forest,coniferous forest and sub-alpine shrubs. We analyzed the soil acidobacterial community composition,elevational patterns and the relationship between Acidobacteria subdivisions and soil enzyme activities by using the 16 S rRNA meta-sequencing technique and multivariate statistical analysis. The result found that 19 known subdivisions as well as an unclassified phylotype were presented in these forest sites,and Subdivision 6 has the highest number of detectable operational taxonomic units(OTUs). A significant single peak distribution pattern(P 0.05) between the OTU number and the elevation was observed. The Jaccard and Bray–Curtis index analysis showed that the soil Acidobacteria compositional similarity significantly decreased(P 0.01) with the increase in elevation distance. Mantel test analysis showed the most of the soil Acidobacteria subdivisions had the significant relationship(P 0.01) with different soil enzymes. Therefore,soil Acidobacteria may be involved in different ecosystem functions in global elemental cycles. Partial Mantel tests and CCA analysis showed that soil pH,soil temperature and plant diversity may be the key factors in shaping the soil Acidobacterial community structure.     

Characterization of extracellular polymeric substances in the biofilms of typical bacteria by the sulfur K-edge XANES spectroscopy

A combined approach of physicochemical extraction and sulfur K-edge X-ray absorption near-edge structure（XANES） spectroscopy was applied to characterize the extracellular polymeric substances（EPS） of typical bacterial biofilms in this study. Physicochemical analysis showed variation of the contents of DNA,polysaccharide and protein in different fractions of EPS in different mediums. The sulfur K-edge XANES analysis yielded a variety of spectra.Spectral fitting of the XANES spectra utilizing a large set of model compounds showed that there was more reduced sulfur in both LB-EPS（loosely bound EPS） and TB-EPS（tightly bound EPS） of all the biofilms in LB medium than in R2 A medium. More oxidized sulfur was identified in LB-EPS than that in TB-EPS,suggesting different niches and physiological heterogeneity in the biofilms. Our results suggested that the sulfur K-edge XANES can be a useful tool to analyze the sulfur speciation in EPS of biofilms.     

电子受体类型对聚磷污泥胞外聚合物的影响研究

采用3种电子受体(硝氮、亚硝氮、氧),在SBR反应器中分别驯化了具有稳定除磷能力的聚磷污泥。对比研究了不同聚磷污泥胞外聚合物(EPS)的组分与含量,结合三维荧光光谱对EPS中有机物质进行分析。研究表明,以硝氮、亚硝氮为电子受体的聚磷污泥EPS平均浓度较为接近,分别为115.6mg/gVSS、133.5 mg/gVSS,由厌氧段至缺氧段,EPS总浓度有所下降。以氧为电子受体聚磷污泥的EPS平均浓度相对较高,为194.5 mg/gVSS,由厌氧段至好氧段,EPS总浓度略有升高。不同聚磷污泥的EPS组分均以胞外蛋白为主,占EPS总量的53%~65%,多糖与核酸占EPS总量28%~34%、5%~7%。三维荧光光谱显示3种聚磷污泥EPS均具有芳香结构蛋白质、可溶性代谢物的荧光峰,另外,以硝氮为电子受体的聚磷污泥还具有明显的腐殖酸类物质的荧光峰。研究结果表明电子受体对聚磷污泥EPS的组分、浓度、有机质类别有一定的影响。     

石油污染对海水中细菌数量和酶活性的影响

在实验室条件下,研究了不同浓度(0,0.05 mg/L,0.15 mg/L,0.45 mg/L和1.35 mg/L)的石油,对海水中细菌数量和酶活性在短期(96 h)和较长期(28 d)内的影响,结果表明:(1)海水中的细菌数量随着石油污染时间和浓度的不同,表现出不同的变化特征。可培养异养菌数在96 h内,所有实验组均呈现先升高后下降的趋势,其后低浓度实验组保持缓慢上升至7 d后数值维持稳定,但最高浓度组(1.35 mg/L)海水中的异养细菌数在7 d后明显下降,之后逐渐上升。实验持续28 d时,各组间差别不显著(P0.05)。在培养24 h时,各实验组细菌总数有明显增长,且随石油浓度增加而升高,随后其变化没有明显规律,但培养7 d后,较低浓度组细菌总数一直高于对照组,而高浓度组(1.35 mg/L)细菌总数一直低于对照组。(2)石油污染对海水中细菌脱氢酶和超氧化物歧化酶(SOD)具有诱导作用。细菌脱氢酶活性和SOD活性分别在培养72 h和24 h时显著升高,且随着石油浓度增加,其活性上升。经过长时间培养后,细菌酶活恢复正常,与对照组没有显著差别。     

不同浓度Cd~(2+)对酿酒酵母的毒害作用研究

以"模式生物"——酿酒酵母(Saccharomyces cerevisiae)为载体,通过测定酵母细胞的生物量、活细胞/死细胞的计数比值以及胞外聚合物(EPS)的含量,研究了不同浓度的Cd2+(Cd2+浓度分别为0.0mg/L、0.1mg/L、0.5mg/L、1.0mg/L、2.0mg/L、8.0mg/L)对酿酒酵母的毒害作用。试验结果表明:当Cd2+浓度小于或等于0.1mg/L时,活细胞/死细胞的计数比值随着时间的推移分别降低了7.86和8.37,在0.5 mg/L、1.0 mg/L和2.0mg/L的Cd2+浓度下,活细胞/死细胞的计数比值则分别上升了3.35、5.64和0.89,8.0mg/L浓度的Cd2+则会完全抑制酵母细胞的增殖,活细胞/死细胞的计数比值波动在0.86~1.11之间;同时,随着Cd2+浓度的上升,酵母细胞的生物量会逐渐减少,0.1mg/L浓度的Cd2+对酵母细胞生物量无明显影响,0.5~8.0mg/L浓度的Cd2+可使酵母细胞生物量比不含Cd2+的培养基分别下降了7.01%、35.14%、59.89%和96.86%;此外,0.5 mg/L和1.0mg/L浓度的Cd2+能显著刺激酵母细胞分泌更多的EPS,增幅分别为20.26%和7.54%。上述结果表明酵母细胞在一定的Cd2+浓度下可以通过提高EPS的产量来抵御Cd2+对细胞的毒性作用。     

中亚热带马尾松林凋落物分解过程中的微生物与酶活性动态

采用凋落物原位分解法,研究了中亚热带马尾松林中马尾松、槲栎凋落叶单独及混合(自然质量比8∶2)分解过程中的微生物数量与酶活性动态.结果表明:①3类凋落物的年分解常数K的大小为:混合凋落物(0.94)>槲栎凋落物(0.86)>马尾松凋落物(0.67);②3类凋落物真菌数量和微生物数量均在夏季(135 d时)达到最大值,而此时细菌和放线菌数量最低;③3类凋落物纤维素酶活性、酸性磷酸酶活性均与凋落物干重剩余率呈显著正相关(P<0.05),而马尾松与混合型凋落物中的多酚氧化酶活性同凋落物干重剩余率呈极显著负相关(P<0.01);④微生物数量和多酚氧化酶活性均总体表现为槲栎凋落物>混合凋落物>马尾松凋落物,酸性磷酸酶活性多表现为槲栎凋落物最低,与分解常数K排列有一定的差异,说明凋落物分解是微生物和多种酶共同作用的结果.整体研究表明,凋落物质量和季节气候的差异显著影响微生物群落及其调控的生态过程,与纯马尾松凋落叶相比,针阔混合使微生物数量和多酚氧化酶活性显著提高,这可能是导致分解加快的重要原因.     

曝气膜生物反应器运行过程中污泥活性特征变化及其对膜污染的影响

研究了曝气膜生物反应器运行过程中活性污泥主要活性特征变化及其对膜污染的影响.通过排出剩余污泥的办法维持活性污泥浓度在4000 mg·L-1左右,并连续运行75 d.运行期间,每日检测活性污泥的各项性质指标便于反映污泥特性的变化.结果表明,随着反应器运行时间的延长,污泥脱氢酶活性逐渐增加,其对反应器的运行有着两方面的作用,一方面会强化微生物对污染物的去除,但另一方面则导致了胞外聚合物的增加,并加速膜污染.而污泥表观产率则随着运行时间的延长先增加后有所减少,其粒径逐渐减小,且胞外聚合物呈现增加的趋势,总的出水水质情况逐渐提高,与此同时,反应器内原生动物及后生动物在运行前期较少,而在后期大量出现.膜污染分析结果表明运行后期膜污染速度明显加快,其原因在于:污泥粒径的减小以及胞外聚合物的增加导致细小颗粒及胞外聚合物堵塞或在膜表面沉积数量增加.     

胞外聚合物磷酸盐形态对生物除磷过程的影响研究

以不同DO条件下污泥龄(SRT)分别为10 d和30 d的两组实验室A/O-SBR反应器活性污泥为研究对象,探讨了胞外聚合物(EPS)磷酸盐形态对生物除磷过程的影响.结果表明,污泥絮体中的磷主要分布于EPS中,PO3-4-P和聚磷酸盐(Poly-P,包括低分子量聚磷酸盐LMW PolyP和高分子量聚磷酸盐HMW Poly-P)是EPS磷的主要形态;EPS对生物除磷的影响明显大于细菌细胞,EPS磷的厌氧降低量和好氧升高量为胞内磷变化量的2.8~6.4倍.EPS中的LMW Poly-P和HMW Poly-P含量均表现厌氧降低和好氧升高的变化规律;对于相同SRT的污泥,中DO(2.5~3.5 mg·L-1)条件较低DO(0.7~1.0 mg·L-1)条件下EPS的LMW Poly-P和HMW Poly-P有更大的厌氧降低量和好氧升高量,对应着更明显的生物除磷过程,说明EPS不仅是生物除磷过程的中转站,而且参与了生物聚磷过程.