Aerobic degradation of methyl tert-butyl ether by a Proteobacteria strain in a closed culture system

The contamination of methyl ten-butyl ether （MTBE） in underground waters has become a widely concerned problem all over the world. In this study, a novel dosed culture system with oxygen supplied by H2O2 was introduced for MTBE aerobic biodegradation. After 7 d, almost all MTBE was degraded by a pure culture, a member of β-Proteobacteria named as PMI, in a closed system with oxygen supply, while only 40% MTBE was degraded in one without oxygen supply. Dissolved oxygen （DO） levels of the broth in closed systems respectively with and without H2O2 were about 5-6 and 4 mg/L. Higher DO may improve the activity of monooxygemase, which is the key enzyme of metabolic pathway from MTBE to tert-butyl alcohol and finally to CO2, and may result in the increase of the degrading activity of PM1 cell. The purge and trap GC-MS result of the broth in closed systems showed that tea-butyl alcohol, isopronol and acetone were the main intermediate products.     

利用泰乐菌素菌渣生产复合酶制剂及效果研究

发酵法生产抗生素过程中会产生大量废弃菌渣,这些菌渣因有机质含量高,堆放极易发生霉变,给当地环境造成危害.文章采用固体发酵法,以微生物降解处理后的泰乐菌素菌渣为主要原料,生产复合酶制剂,并对其在肉鸡上的使用效果进行研究.结果表明:在水含量50％～60％、干物质中麸皮含量不低于40％的泰乐菌素菌渣培养基中,接入5％～10％...     

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.     

纳米银与石墨烯对土壤微生物及土壤酶的影响

采用室内暗培养试验分别探究了纳米银与石墨烯对土壤微生物及土壤酶的不同影响.将不同剂量的纳米银(0、10、100、150 mg·kg~(-1))与高纯石墨烯(0、10、100、1000 mg·kg~(-1))分别与等量棕壤充分混匀,然后进行暗培养.在第3、7、15、30和60 d时取样,测定土壤脲酶、土壤碱性磷酸酶、土壤脱氢酶和土壤过氧化氢酶的活性及土壤细菌、真菌和放线菌的数量,并在培养期间测定土壤呼吸速率及CO2累积量.结果表明,所有纳米银处理均抑制土壤的呼吸作用,并且剂量越高,抑制作用越明显;而石墨烯处理未对土壤呼吸产生显著影响.10 mg·kg~(-1)纳米银处理下,土壤真菌数量在整个培养期内均显著低于对照,土壤细菌在第60 d时也被显著抑制,但土壤放线菌数量无变化;与对照相比,100和150 mg·kg~(-1)的纳米银处理显著降低了土壤细菌、真菌、放线菌的数量.10和100 mg·kg~(-1)的石墨烯处理下,土壤细菌、真菌、放线菌数量则均无显著变化.1000 mg·kg~(-1)的石墨烯显著增加了土壤中细菌与真菌的数量,却对土壤放线菌数量无影响.纳米银处理显著抑制土壤脲酶、脱氢酶活性,却对土壤过氧化氢酶与磷酸酶活性基本无影响.10和100 mg·kg~(-1)石墨烯处理对土壤脲酶有一定的促进作用,1000 mg·kg~(-1)石墨烯处理对土壤过氧化氢酶和脱氢酶有一定的促进作用,而不同剂量的石墨烯在培养后期均对碱性磷酸酶产生抑制作用.总体来说,纳米银在一定程度上对土壤酶及土壤微生物结构产生了负面影响,而石墨烯对土壤酶及土壤微生物结构的影响不明显.     

纤维素降解菌Arthrobacter oryzae HW-17的纤维素降解特性及纤维素酶学性质

采用单一碳源选择性培养基和纤维素平板水解圈法筛选到一株具有较强纤维素分解能力的菌株Arthrobacter oryzae HW-17.此外,高通量测序发现,不同驯化条件下微生物群落结构有明显差异,低温条件下优势属为类芽孢杆菌属(Paenibacillus)和伯克氏菌属(Burkholderia).本文同时对菌株Arthrobacter oryzae HW-17的微生物特性和纤维素降解特性进行了初步研究,结果发现,KNO_3、30或35℃、pH=7分别为菌株产纤维素酶的最佳氮源、温度和pH.菌株HW-17的最高纤维素酶活为18.55 U·m L~(-1),且对磨碎加工处理的纤维素样品和含鸡粪的纤维素混合样品有更好的降解效果.此外,菌株HW-17产生的纤维素酶在中温(≤50℃)和偏酸性(pH=5~7)条件下能保持较高的酶活.Cu(Ⅱ)和Pb(Ⅱ)等金属离子能够抑制该酶的活性.     

Effect of some herbicides on activities of microorganisms and enzymes in soil

Abstract

Laboratory tests were conducted with eight herbicides, atrazine, butylate, ethalfluralin, imazethapyr, linuron, metolachlor, metribuzin and trifluralin, applied to a loamy sand at rate of 10 μg/g to determine if these materials caused any serious effects on microbial and enzymatic activities related to soil fertility. Some herbicides showed an effect on bacteria and fungi for the first week of incubation, but, subsequently, the populations returned to levels similar to those obtained in the controls. After several herbicide treatments there appeared to cause a slight depression of nitrification. Sulfur oxidation was better than that obtained with untreated soil in all treatments. Oxygen consumption was increased significantly after 96 hr incubation with atrazine. The soil dehydrogenase and amylase activities were inhibited by ethalfluralin treatment respectively for 1 wk and 1 day, and p‐nitrophenol liberation was inhibited for 2 hrs by all herbicide treatments. Results indicated that the herbicidal treatments at the level tested were not drastic enough to be considered deleterious to soil microbial and enzymatic activities which are important to soil fertility.     

Effect of tracer dyes on initial deposits and persistence of bacillus thuringiensis subsp. kurstaki toxin after application of two commercial formulations onto spruce trees

Abstract

The effect of two tracer dyes [Erio Acid Red (EAR) and Acid Black 48 (AB‐48)] on initial deposits and persistence of Bacillus thuringiensis subsp. kurstaki (Btk) toxin (delta‐endotoxin) was studied after spraying two commercial formulations, Foray® 48B and Foray® 76B, over potted white spruce [Picea glauca (Moench) Voss] seedlings, at a dosage rate of 30 billion international units (BIU) per ha. Spray was applied using a spinning disc atomizer calibrated to deliver droplet sizes similar to those utilized in ultra‐low‐volume (ULV) treatments in operational insect control programs. The sprayed seedlings were left outdoors at the Sault Ste. Marie laboratory for 18 days under natural conditions of sunlight, wind and rainfall. Initial deposits and persistence of delta‐endotoxin protein in spruce foliage were determined by immunoassay [enzyme linked immunosorbent assay (ELISA)] quantification of the delta‐endotoxin. The total protein (inactive plus active) and delta‐endotoxin (active protein) concentrations in the two formulations were determined by a gravimetric procedure and by ELISA respectively.

The initial deposit levels of the toxin on foliage were not markedly affected by the addition of either of the two tracer dyes, and showed only a narrow range of 1521 to 1625 ng/g foliage (fresh weight) for Foray 48B, and 1789 to 2056 ng/g for Foray 76B. However, the persistence of the toxin was significantly influenced by the presence of the dyes. The toxin persisted in foliage only for 7 d post‐spray When the EAR dye was added to Foray 48B, compared to 10 d when no dye was added. The average half‐life (DT₅₀) of disappearance was 17.4 h for Foray 48B with EAR, and 20.9 h when no dye was present. In contrast, the situation was reversed in Foray 76B, since the duration of persistence was 10 d when EAR was added to Foray 76B, compared to 7 d when no dye was added. The average DT₅₀ was 27.9 h for Foray 76B with EAR, and 22.2 h without the dye. Persistence was the longest (14 d) when the AB‐48 dye was added to Foray 76B, and the DT₅₀ was 44.9 h.     

Progress in the genetic manipulation of crops aimed at changing starch structure and increasing starch accumulation

The starch content and its composition have important consequences for the yield of the harvested crop and the materials extracted from it. The functional properties of the foods or other processed materials derived from these crops are also affected by the structure and composition of the starch. Recently, genetic engineering has been used to produce plants with an elevated starch content, achieved by transforming the plant with a mutated bacterial gene coding for an ADPglucose pyrophosphorylase that is active in the presence of metabolites which inhibit the plant enzyme. Besides the practical implications of these results, this experiment provided direct evidence for the regulatory role of the ADPglucose pyrophosphorylase in starch synthesis. Other bacterial enzymes, such as glycogen synthase and branching enzyme, could be introduced in order to modify starch structure. However, a more elegant (but longer-term) approach would be to learn enough about the structure-function relationships of the plant enzymes so that the product of their action could be changed. To achieve this objective, much more will have to be learned about the enzymes involved in the biosynthesis of starch than is presently known. Here, the basic properties of starch and the current research approaches to understanding its biosynthesis are described, together with a perspective of how genetic manipulation of starch structure may be achieved.Paper presented at the Bio/Environmentally Degradable Polymer Society—Third National Meeting, June 6–8, 1994, Boston, Massachusetts.     

Purification and characterization of extracellular poly(3-hydroxybutyrate) depolymerases

Five extracellular PHB depolymerases of bacteria isolated from various sources were purified to electrophoretic homogeneity and compared with known extracellular PHB depolymerase fromAlcaligenes faecalis T1. The molecular mass of these enzymes were all around 40–50 kDa. Nonionic detergent, diisopropylfluorophosphate and dithiothreitol inhibited the PHB depolymerase activity of all these enzymes. Trypsin abolished PHB depolymerase activity, but not theD-3-hydroxybutyric acid dimer hydrolase activity of all the enzymes. These results showed that the basic properties of these PHB depolymerases resemble those of theA. faecalis T1 enzyme. Analysis ofN-terminal amino acid sequence of the purified enzymes revealed that these enzymes includingA. faecalis T1 enzyme fall into three groups.     

The Role Played by Acetyl Group Distribution Patterns in Substituted Starch Toward Enzymatic De-acetylation and Chain Fragmentation

The nature and distribution of the acetylated groups were evaluated by ¹³C-NMR and ¹H-NMR. The starch substrate with a DS of 1.5 comprises only two patterns: -(14)-d-glucopyranose and 2,3,6-tri-O-acetyl--(14)-d-glucopyranose. The starch with a DS of 3.0 also comprises two patterns: 2,3,4,6-tetra-O-acetyl--(14)-d-glucopyranose and 2,3,6-tri-O-acetyl--(14)-d-glucopyranose; whereas starch (DS = 1.9) contains 4 patterns: 2,3,6-tri-O-acetyl--(14)-d-glucopyranose, 2,3,4,6-tetra-O-acetyl--(14)-d-glucopyranose terminal, 2,6-di-O-acetyl--(14)-d-glucopyranose, and 3,6-di-O-acetyl--(14)-d-glucopyranose. Using esterase from Viscozyme, it has been possible to hydrolyze up to 7% of the DS 3.0 starch. An -amylase (Fungamyl 800) was then added to these acetylesterases. With a 2.4 FAU/mL fraction of -amylase and 2.4 U/mL from the Viscozyme's acetylesterase, 28% of the acetylated end groups were hydrolyzed for the starch substrates with DS 3.0. Moreover, a synergic action between -amylase and acetylesterase was noticed, allowing fragmentation of 32% for DS 1.5, 30% for DS 1.9, and 11% for DS 3.0.