A Novel Gene Encoding an Enzyme That Degrades a Polysaccharide from the Sheath of Sphaerotilus natans

A gene encoding an enzyme that is able to depolymerize the basic polysaccharide prepared from the sheath of Sphaerotilus natans was identified in a sheath-degrading bacterium, Paenibacillus koleovorans. The gene was constructed from 2217 bp coding for 738 amino acids, including the signal sequence of 34 amino acids. No closely related protein or gene was indicated by a homology search. The gene was expressed in Escherichia coli as a glutathione S-transferase fusion protein. The fusion protein depolymerized the sheath polysaccharide into an oligosaccharide, introducing an unsaturated sugar residue, suggesting that the gene codes for a polysaccharide lyase acting on a basic polysaccharide.     

Enantioseparation and enantioselective behavior of trichlorfon enantiomers in sediments

Trichlorfon (TF), an organophosphorus insecticide, has been widely used in seawater aquaculture; it is easily degraded to the highly toxic insecticide, dichlorvos (DDVP). In this study, the enantioseparation of TF enantiomers, as well as their degradation behavior and product (DDVP) formation in mariculture pond sediments, was investigated. The results show that both TF enantiomers degrade into DDVP, which is the main degradation product. Furthermore, S ‐(+)‐TF is preferentially degraded under natural conditions, suggesting that TF enantiomers degrade enantioselectively. Nevertheless, the degradation behavior of TF enantiomers is not enantiospecific under sterile conditions. The formation of DDVP and the enantiospecific degradation of TF enantiomers are attributed to the activities of microbes present in the sediments.     

Decrease of proteolytic degradation of recombinant hirudin produced by Pichia pastoris by controlling the specific growth rate

The effects of the specific growth rate and methanol concentration on the degradation of hirudin produced by recombinant Pichia pastoris were investigated. When a methanol-limited state and the specific growth rate of 0.02 h^–1 were maintained during the fermentation, a minimum degradation of hirudin and a maximum specific hirudin production rate were achieved. By this strategy, the production of intact recombinant hirudin Hir65 reached 0.7 g l^–1 in fed-batch fermentation. Its proportion was 35% to all forms of hirudin.     

Cellulose depolymerization to glucose and other water soluble polysaccharides by shear deformation and high pressure treatment

The simultaneous action of shear deformation and high pressure (SDHP) creates changes in the structure of wood and its main components (cellulose, hemicelluloses, lignin). The formation of water and alkali soluble polysaccharides under SDHP action, proceeds in seconds in the solid state, without the use of any reagents and solvents. Therefore, SDHP seems to be a technologically safe method and friendly to the environment. The amorphization of cellulose crystallites and depolymerization of cellulose chains were observed under a wide range of pressures (1–6 GPa), both for cellulose samples and the cellulose part of wood. Similar depolymerization occurs in the hemicellulose part of wood. The decomposition of polysaccharides under SDHP causes the formation of the water soluble part, whose content increases with pressure and the applied shear deformation. A maximum solubility of 40% and 55% was registered at 6 GPa following treatment of cellulose and birch wood samples. A higher output in the case of wood can be explained by a specific role of lignin under SDHP, which acts as a ‘grinding stone’ during cellulose and hemicelluloses destruction. As shown by high-performance size exclusion chromatography, the water soluble fraction obtained from cellulose contained glucose (2.6%), cellobiose (9.6%), cellotriose (16.6%) and other higher water soluble oligomers (71%). Almost complete dissolution (98%) of the treated cellulose sample can be achieved by extraction with 10% NaOH solution. The SDHP treated birch wood was subjected to submerged fermentation (with Trichoderma viride), and a 13% output of proteins was obtained. In this case, the water soluble part played the role of the so called ‘start sugars’. Abbreviations: ASF, alkali soluble fraction; DP, degree of polymerization; EC, energy consumption; HP, high pressure; LMWS, low molecular weight sugars; MC, moisture content; MCC, microcrystalline cellulose; SD, shear deformation, SDHP, shear deformation under high pressure; SS, shear strength; WSF, water soluble fraction This revised version was published online in November 2006 with corrections to the Cover Date.     

An Improved Method to Obtain High Molecular Weight DNA from Purified Micro- and Macronuclei of Tetrahymena thermophila

An improved method to obtain high molecular weight DNA from purified macro- and micronuclei of Tetrahymena thermophila is described. Micro- and macronuclear DNA obtained using previously described protocols was degraded and not suitable for the cloning of large (> 100 kb) DNA fragments. Based on the data reported here, we propose that DNA degradation is mainly due to nuclease activity; some micronuclear DNA degradation is due to mechanical shearing as a result of extended periods of blending. We have made modifications to reduce nuclease degradation by minimizing cell lysis, by the early addition of EDTA and by increasing the EDTA concentration (23 mM). To reduce mechanical shearing, cell and nuclear suspensions were blended for shorter periods. High molecular weight micro- and macronuclear DNA was obtained using the new protocol.     

The influence of moisture on microbial transport, survival and 2,4-D biodegradation with a genetically marked Burkholderia cepacia in unsaturated soil columns

The influence of moisture on the survival, movement anddegradation activity of a 2,4-D degrading bacterium,Burkholderia cepacia strain BRI6001L, geneticallyengineered to contain bioluminescent and lactoseutilization genes, was studied in unsaturated soil columns.The distance traveled by BRI6001L was dependent on theclay content of the soil, higher clay contents beingresponsible for higher filtration coefficients. Long termsurvival, in excess of one year, was attributed to strainBRI6001L's ability to survive dry conditions. Changes inthe 2,4-D biodegradation rate showed a better correlationwith the BRI6001L population density than with the totalviable bacterial population. At moisture levels betweenfield capacity and 40% moisture (– 33 kPa to –100 kPa)2,4-D degradation was attributed mainly to BRI6001L. Atmoisture levels between 6 and 15%, 2,4-D disappearancewas attributed to the indigenous microbial population,with no degradation occurring at moisture levels below6%. Returning the moisture to above 40% led to anincrease of 4 orders of magnitude in the BRI6001Lpopulation density and to a 10-fold increase in the 2,4-Ddegradation rate. The ability to monitor a specificmicrobial population using reporter genes hasdemonstrated the importance of controlling moisturelevels for maximizing biodegradation rates in unsaturatedsoil environments.     

棘冠海星暴发及其对珊瑚礁的生态影响研究进展

棘冠海星的反复暴发是导致印度-太平洋区域珊瑚礁生态系统退化的最主要原因之一。然而，我国对棘冠海星的研究非常有限。本文综述了国内外关于棘冠海星及其暴发的生态影响和应对策略的研究进展，得出以下主要结论：1）雌性棘冠海星个体每年产卵数量高达50万-2亿个，环境因素变化只要导致幼虫和幼体存活率的轻微提高，成体就将得到大量补充；2）棘冠海星暴发的阈值为1000-1500个/km²，暴发周期为10-27 a，每次暴发持续1-10 a，最终可能以"种群集体感染疾病而崩溃"结束；3）棘冠海星暴发对印度洋及太平洋东部和北部珊瑚礁的破坏性非常小，却直接导致太平洋的西部和南部珊瑚礁90%以上的珊瑚死亡，并通过改变珊瑚群落组成、减少珊瑚和鱼类多样性而对珊瑚礁产生间接影响；4）关于棘冠海星暴发原因的假说中"陆地营养物质输入假说"和"捕食者过度捕捞假说"得到了最普遍的认可，但都不能解释所有的暴发事件；5）应对棘冠海星暴发的主要策略有改善水质、设立保护区、投放天敌和人工清理等，其中人工清理是最直接有效的策略，但迄今并没有发现可长期抑制棘冠海星暴发的方法。因此，急需加强对棘冠海星的深入研究，探查其暴发的根本原因和对策。