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.     

Ammonium,Nitrate and Nitrite Nitrogen and Nitrate Reductase Enzyme Activity in Groundnut (Arachis hypogaea L.) Fields After Diazinon,Imidacloprid and Lindane Treatments

Impacts of diazinon (O,O-diethyl O-2-isopropyl-6-methylpyrimidin-4-yl phosphorothioate), imidacloprid [1-(6-chloro-3-pyridylmethyl)-N-nitroimidazolidin-2-ylideneamine] and lindane (1,2,3,4,5.6-hexachlorocyclohexane) treatments on ammonium, nitrate, and nitrite nitrogen and nitrate reductase enzyme activities were determined in groundnut (Arachis hypogaea L.) field for three consecutive years (1997 to 1999). Diazinon was applied for both seed- and soil-treatments but imidacloprid and lindane were used for seed treatments only at recommended rates. Diazinon residues persisted for 60 days in both the cases. Average half-lives (t_1/2) of diazinon were found 29.3 and 34.8 days respectively in seed and soil treatments. In diazinon seed treatment, NH₄ ⁺, NO₃ ^?, and NO₂ ^? nitrogen and nitrate reductase activity were not affected. Whereas, diazinon soil treatment indicated significant increase in NH₄ ⁺-N in a 1-day sample, which continued until 90 days. Some declines in NO₃ ^?N were found from 15 to 60 days. Along with this decline, significant increases in NO₂ ^?N and nitrate reductase activity were found between 1 and 30 days. Imidacloprid and lindane persisted for 90 and 120 days with average half-lives (t_1/2) of 40.9 and 53.3 days, respectively. Within 90 days, imidacloprid residues lost by 73.17% to 82.49% while such losses for lindane residues were found 78.19% to 79.86 % within 120 days. In imidacloprid seed-treated field, stimulation of NO₃ ^?N and the decline in NH₄ ⁺NO₂ ^?-N and nitrate reductase enzyme activity were observed between 15 to 90 days. However, lindane seed treatment indicated significant increases in NH₄ ⁺-N, NO₂ ^?-N and nitrate reductase activity and some adverse effects on NO₃ ^?N between 15 and 90 days.     

Spore inhibition-based enzyme substrate assay for monitoring of aflatoxin M1 in milk

A spore germination-based concept and its transformation into a field level prototype for monitoring aflatoxin M₁ (AFM₁) in milk was developed. Initially, 15 strains of Bacillus spp. procured from different culture collection were screened for AFM₁ sensitivity using spot assay and marker strain showing inhibition at 0.5 ppb was selected based upon maximum zone of inhibition. The selected strain B. megaterium 2949 was further screened for different enzymes activities and subsequently its spores were produced to an extent of 73.13% ± 3.197% in newly developed sporulation medium containing beef extract (0.0075% ± 0.0004%), yeast extract (0.015% ± 0.001%), peptone (0.0375% ± 0.0016%), and sodium chloride (0.0375% ± 0.0018%). A spore germination-based concept/ assay was optimized by immobilizing spores in eppendorf with pretreated milk (80°C/15 min) containing germinant and chromogenic substrate followed by incubation at 37°C. The appearance of sky blue color within real time of 45 min indicated spores germination and release of specific marker enzyme such as acetyl esterase and its specific action on chromogenic substrate which demonstrates absence of AFM₁ in milk. However, if there was no color change, presence of AFM₁ at 0.5 ppb MRL was denoted by Codex. The developed concept on AFM₁ detection was validated and a correlation of 0.97 was established with AOAC approved Charm 6602 and ELISA at Codex MRL with minimal false positive and negative results. The cost effective test has potential application in dairy farms, manufacturing, and R&D units for routine monitoring of AFM₁ in milk.     

藻-菌颗粒污泥中微生物胞外多糖特性研究

藻-菌颗粒污泥（Microalgal-bacterial granular sludge,MBGS）工艺在城市污水生物处理领域引起了越来越多的研究兴趣.胞外多糖（Polysaccharides,PS）是胞外聚合物（Extracellular polymeric substances,EPS）的重要组分,是维持颗粒稳定性的关键物质之一.研究了MBGS中PS的凝胶性能,以及松散结合的胞外聚合物（Loosely bound EPS,LB-EPS）和紧密结合的胞外聚合物（Tightly bound EPS,TB-EPS）中PS的含量、分布及单糖组成,并探究了MBGS的EPS中PS可能的微生物来源.结果表明,MBGS的PS具有凝胶性,主要分布在TB-EPS中,含量为 （40.06±2.30） mg·g^-1,其中半乳糖、葡萄糖和鼠李糖为主要单糖组分,而果糖和古罗糖醛酸未被检出.Thauera、Hydrogenophaga、Azoarcus和Acutodesmus是PS的主要微生物潜在来源.本文研究结果阐释了MBGS PS的基本特性,为MBGS工艺的稳定运行提供了理论基础.     

典型氢氧化细菌固碳特性种间差异及胞外有机物对固碳过程影响

化能自养细菌对全球CO₂固定具有重要的意义.研究了4种典型氢氧化细菌（HOB,分别是Alcaligenes hydrogenophilus DSM 2625、Pelomonas saccharophila DSM 654、Variovorax paradoxus DSM 30034和Acidovorax facilis DSM 649）的固碳特性及其种间差异性,并探究了胞外游离有机碳（EFOC）对不同种类HOB固碳效率差异性的影响.结果表明：①不同HOB自养培养过程中表观固碳量随时间呈显著差异性,其中,DSM 2625生长速度较快,其平均固碳量分别是DSM 654、DSM 649和DSM 30034的6.30、8.76和7.02倍.②不同HOB之间cbbL基因转录量与 表观固碳量的相关系数为0.980 （p<0.05）,cbbL基因转录量是造成不同菌种之间表观固碳效率差异的关键因素.③HOB细胞蛋白质含量与cbbL基因丰度和转录量之间也存在显著正相关,相关系数分别为0.999（p<0.01）和0.976（p<0.05）,即化能自养细菌CO₂同化途径所固定的有机碳进一步参与细胞蛋白质的合成,进而影响其生长速度和表观固碳效率.④HOB在自养培养过程中产生的EFOC对cbbL基因转录效率产生反馈抑制作用,且不同HOB之间,EFOC/TOC比例越低,表观固碳效率越高.     

三峡库区消落带落羽杉人工林土壤细菌群落结构多样性及动态变化

消落带是连接陆地与水域的交错地带,具有重要的生态功能,其适生木本植物的种植对消落带土壤的生物地球化学循环起着重要的作用.为探究适生植物对三峡库区消落带环境的适应机制,采用高通量测序技术对消落带退水后适生植物落羽杉(Taxodium distichum)不同生长时期(T1:5月、 T2:7月和T3:9月)的土壤细菌群落组成及多样性进行研究,同时采用PICRUSt2对细菌功能进行预测.结果表明,土壤pH值、硝态氮、铵态氮、土壤蔗糖酶、磷酸酶和脲酶等理化指标随时间变化显著(P<0.05);土壤细菌多样性、丰富度和结构也随时间变化而变化,除Chao1以外,根际土壤细菌的α多样性均表现为：T1>T2>T3,非根际土壤细菌α多样性则表现为：T3>T1>T2. RDA分析表明,影响细菌群落的理化指标主要有土壤pH值、脲酶、铵态氮和硝态氮.所有土壤样本中共检测到细菌60门,其中以变形菌门和酸杆菌门为优势细菌门.根据PICRUSt2预测,代谢是落羽杉土壤细菌群落中普遍存在的基本功能,与C、 N和P有关的各代谢途径存在一定的时间差异.以上研究结果有助于加深对三峡消落带植被修复...     

牡蛎壳粉和石灰改良酸性水稻土对磷有效性、形态和酶活性的影响

我国南方粮食主产区土壤酸化治理是提升耕地地力促进粮食增产的重要内容,目的是探究酸化土壤改良剂牡蛎壳粉和石灰对水稻土酸度、磷有效性和形态以及酶活性和相关功能基因的影响.盆栽试验设计2种改良剂的3个施用量（0.05%、0.10%和0.15%）,以水稻为试验作物.结果表明：①牡蛎壳粉和石灰均显著提高土壤pH,降低土壤交换性酸,且改良效果随着改良剂用量增加而提高,相同用量下石灰对土壤酸性的改良效果高于牡蛎壳粉;② 4种有效磷提取剂提取的土壤有效磷测定结果表明,牡蛎壳粉和石灰显著提高H₂SO₄-P、Bray-1 P和Olsen-P含量,其中Olsen-P含量随改良剂用量的增加而提高.③土壤中不同形态无机磷含量表现为：Fe-P>Al-P>Ca-P,石灰和牡蛎壳粉显著提高Al-P和Fe-P含量,分别提高26.3%~37.4%和7.9%~23.7%;0.15%改良剂显著增加Ca-P含量;④石灰和牡蛎壳粉对土壤DHA、ALP、IPP活性和phoD基因拷贝数具有提高作用,而对土壤ACP活性、phoC和pqqC拷贝数表现为降低;⑤改良剂（0.10%和0.15%）显著提高水稻产量,0.15%用量的石灰和牡蛎壳粉分别提高水稻产量34.2%和46.8%,但对秸秆生物量无显著影响.相关分析表明,土壤无机磷和有效磷含量与pH和ALP活性呈显著正相关,改良剂通过消除土壤酸化和提高土壤磷酸酶活性来改善土壤磷营养状况,这对促进作物产量具有积极作用.     

生物炭施用对黄壤土壤养分及酶活性的影响

生物炭因其具有特殊的理化性质,作为土壤改良剂或调理剂被广泛应用于改善土壤质量;土壤养分与土壤酶活性是表征土壤质量化学性质和生物学性质的重要指标.采用大田试验,研究生物炭不同施用水平：0（CK）、5（B5）、10（B10）、20（B20）和50（B50） t·hm^-2对黄壤养分和土壤酶活性的影响,运用结构方程模型（SEM）定量分析生物炭处理对土壤养分和酶活性的直接或间接影响及其作用大小.结果表明,生物炭显著增加土壤pH值、电导率、有机碳、碱解氮、有效磷和速效钾（P<0.05）;随生物炭施用量的增加,土壤过氧化氢酶和脲酶活性先增加后降低,磷酸酶和蔗糖酶活性增加（P<0.05）;B10处理下,土壤过氧化氢酶、脲酶和磷酸酶的活性均达到最大值,蔗糖酶活性也相对较高.随生物炭作用时间增加,土壤pH值、碱解氮、有效磷和速效钾含量均增加,而电导率和有机碳与之相反;过氧化氢酶活性降低,脲酶和磷酸酶活性升高,蔗糖酶活性无明显变化规律.SEM结果显示,生物炭施用对过氧化氢酶具有直接的负效应;通过提升pH、电导率、有机碳和碱解氮含量间接影响过氧化氢酶活性,通过提升pH和电导率间接促进蔗糖酶活性,通过提高电导率以及碱解氮和有效磷含量间接增加磷酸酶活性.综上,生物炭施用量及作用时间显著影响土壤养分含量,进而间接作用于土壤酶活性;酸性黄壤施用10 t·hm^-2的生物炭较为适宜.     

甲苯对土壤芳基硫酸酯酶活性的影响

甲苯作为土壤酶测定的前处理试剂及许多有机污染物在环境中降解的中间产物,有关其对土壤酶的影响研究在土壤酶学和环境科学领域具有重要意义.因此,采用模拟方法,较系统地研究了不同剂量甲苯和处理时间下芳基硫酸酯酶活性的变化规律.结果表明,甲苯对纯酶具有明显的抑制作用,酶活性降幅最大达到45.5%;灭菌土壤对溶液中的纯酶有很强的吸附能力;极微量的甲苯即可完成对土壤中酶活性的激活作用,酶活性增幅为109%～298%;随着甲苯剂量的增加,土壤酶活性的变化幅度逐渐趋缓,并可用Langmuir模型较好地拟合,由此获得了最大表观酶活性Umax,发现其与土壤性质显著相关,说明甲苯主要是通过杀死土壤中的微生物来影响土壤酶活性的.此外,初步探讨了不同土壤中胞外酶量与胞内酶量的关系,发现在供试土样中土壤芳香硫酸酯酶胞外酶和胞内酶分别占54.4%和45.6%.本研究可为后续土壤酶测定质量的完善和提高提供依据.     

镉和乙草胺复合污染对玉米生理特性的影响

采用盆栽试验,研究了镉与乙草胺复合污染对玉米幼苗MDA含量和抗氧化酶活性的影响。结果表明,镉单独处理,低浓度下MDA的含量变化不大,当镉浓度大于30 mg/kg时,MDA的含量急剧上升;超氧化物歧化酶SOD、CAT在浓度为30 mg/kg时活性达到最高,之后随浓度的增大,其活性逐渐降低;POD的活性随着镉浓度的增大逐渐升高,且100 mg/kg时达到最高。镉与乙草胺复合处理,低镉浓度下MDA的含量与对照组CK相比差异显著,处理浓度越大,MDA含量越高;SOD、CAT的活性变化趋势与单镉处理时相类似,但变化幅度更加明显;POD的活性则随着浓度的增大继续升高。镉与乙草胺复合处理对玉米幼苗的毒害程度要大于单镉处理,并且施加乙草胺的浓度越大,玉米受毒害程度越深。