鲜木薯抗性淀粉的制备与性质

为了提高木薯抗性淀粉的含量,该研究以鲜木薯湿淀粉为原料,采用压热-酶法制备抗性淀粉,通过单因素试验和响应面分析,获得抗性淀粉的最佳制备条件为：淀粉乳浓度10%、压热时间80 min、压热温度120℃、耐热α-淀粉酶添加量1 U/g、耐热α-淀粉酶作用时间15.75 min,普鲁兰酶添加量0.83 U/g,普鲁兰酶作用时间5.86 h、超声波处理时间2 min。在此条件下抗性淀粉的质量分数是15.48%。电镜试验表明淀粉颗粒经压热-酶法处理后表面形态发生变化;X-射线衍射表明抗性淀粉的结晶类型为B型,结晶度增加;体外消化模拟试验表明：与原淀粉相比,抗性淀粉消化特性降低。该研究可为抗性淀粉的工业化生产和应用提供参考。     

普鲁兰酶氮端氨基酸的截短对其酶学性质的影响

淀粉是丰富的生物质资源,已被广泛地应用到食品、酿酒、医药等各个相关的领域.普鲁兰酶(pullulanase,pulA,EC 3.2.1.41)作为淀粉脱支酶,可以水解淀粉中的α-1,6糖苷键,最大限度地降解淀粉原料,提高淀粉利用率.因此,寻找一条国产化的道路开发我国自己的普鲁兰酶,具有长远的意义.课题组前期从淀粉厂附近的土壤中筛到一株普鲁兰酶的高产菌株变栖克雷伯氏菌(Klebiella variicola)strain 7,克隆获得pulA基因(GenBank登录号:KJ146839.1)后在大肠杆菌(Escherichia coli) BL21 (DE3)中异源表达.为提高该蛋白的表达量和分泌效率,同时改善普鲁兰酶的性质,本研究利用基因工程的手段构建了普鲁兰酶去信号肽突变体M1和氮端31个氨基酸的截短突变体M2,结果显示M1和M2最适温度均为45 ℃,二者的最适pH分别是6.0和5.6;M2的半衰期是37 min,是M1的6.17倍;M2的比酶活是582.204U/mg,是M1的1.6倍.动力学分析显示,以普鲁兰糖为底物时,M2的Vmax、Kcat、Km和Kcat/Km分别为0.001 3μmoL/(mL·s)-1、191.80-1 s.-1、0.30 mg/mL、693.30,与M1相比,M2的底物亲和能力增加,同时催化效率是M1的2倍.本研究通过普鲁兰酶氮端氨基酸的截短,为提高酶的比酶活和半衰期提供了新的方法和思路.     

玉溪醇化烟叶表面细菌酶制剂对烟叶中淀粉和纤维素的降解作用

合理降解烤后烟叶中的淀粉和纤维素含量已成为提高烟叶质量的关键工艺之一。本研究对分离自烤烟叶面的产淀粉酶细菌菌株解淀粉芽孢杆菌(Bacillus amyloliquefaciens A1)和产纤维素酶细菌菌株短小芽孢杆菌(B.pumilus C1)进行发酵培养,提取粗酶液,检测酶活性并应用于初烤烟叶上。通过设计不同酶量、作用时间、相对湿度和温度的方法考察这两种酶制剂对烟叶(Nicotiana tabacum K326)中淀粉和纤维素的降解效果。结果表明,菌株A1主要产α-淀粉酶,酶活力为7×105U/mL,菌株C1以产内切酶为主,酶活力为6×103U/mL;在温度40℃、湿度70%条件下作用96h,用菌株A1所产淀粉酶制剂4×107U/kg处理后的烟叶总糖增加率为12.78%,还原糖增加率为12.03%,菌株C1所产纤维素酶制剂4×105U/kg处理后的烟叶总糖增加率为13.87%,还原糖增加率为18.07%。研究结果提示,利用从烟叶表面分离的产酶菌株进行发酵生产的酶制剂可降解初烤烟叶中淀粉和纤维素,提高还原糖含量,可望在烟叶加工过程中得到应用。     

低温挤压添加酶制剂的玉米粗淀粉的糖化过程试验研究

为了缩短生产玉米糖浆的玉米粗淀粉的糖化时间，改进其糖化过程。通过实验室试验,研究了添加酶制剂的玉米粗淀粉的挤压-液化系统诸参数(套筒温度、玉米粗淀粉含水率、螺杆转速、挤压时耐高温α-淀粉酶添加量、液化时耐高温α-淀粉酶添加量)，对糖液的主要考察指标(糖液的DE值、出品率和过滤速度)的影响规律。研究表明，在本研究的较优系统参数下，糖化12 h糖液的DE值、过滤速度和淀粉出品率分别为95.5%～96.3%、163.2～177.3 L/(m²·s)和94.2%～94.4%。比传统玉米淀粉糖化时间缩短约18 h。     

酶解薯类淀粉适用于电镜观察其颗粒表面及内部结构

为了同步研究淀粉颗粒表面小体和壳层结构,用α-淀粉酶、β-淀粉酶和普鲁兰酶,在室温下单一或复合酶解马铃薯、红薯、木薯淀粉颗粒,用扫描电子显微镜观察酶解颗粒并进行性质测试。研究结果表明,单一酶作用时,只有α-淀粉酶可使3种薯类淀粉显露颗粒表面小体(直径29～73 nm)和壳层结构(厚度150～400 nm);马铃薯淀粉的酶解率(1.1%)远低于其他2种淀粉的(14.1%、16.3%)。马铃薯淀粉表面小体的排列较紧密、壳层结构较致密,决定了其具有较强的抗酶解性和较大的峰值黏度(即膨胀能力)。复合酶作用时,α-淀粉酶复合与其单一作用的效果类似。因此单一α-淀粉酶有限酶解法可以作为淀粉颗粒表面小体和壳层结构的研究方法,酶解条件为:酶浓度80 U/mL,室温下酶解12 h。该研究结果为淀粉类产品在实际加工过程中的品质控制提供了理论基础。     

两种淀粉酶的酶学性质及应用研究

研究了两种耐高温α-淀粉酶和两种真菌α-淀粉酶的酶学性质,确定了最佳酶反应条件.将其应用于淀粉糖生产中,以30%的玉米淀粉为原料,用进口耐高温α-淀粉酶水解至还原糖含量为16.5%,再用日本真菌α-淀粉酶在最佳条件下反应21 h,可得到含纯麦芽糖31.1%、葡萄糖1.7%、糊精2.7%的高麦芽糖浆.     

两种淀粉酶的酶学性质及应用研究

研究了两种耐高温α-淀粉酶和两种真菌α-淀粉酶的酶学性质，确定了最佳酶反应条件。将其应用于淀粉糖生产中，以30%的玉米淀粉为原料，用进口耐高温α-淀粉酶水解至还原糖含量为16.5%，再用日本真菌α-淀粉酶在最佳条件下反应21 h，可得到含纯麦芽糖31.1%、葡萄糖1.7%、糊精2.7%的高麦芽糖浆。     

热硫化氢高温厌氧杆菌的Ttx31基因克隆、表达与功能分析

生物信息学分析发现,热硫化氢高温厌氧杆菌(Thermowmerobacter thermohydrosulfuricus)的α-淀粉酶/普鲁兰糖酶(α-amylase/pullulanase)基因中的一段未知功能序列Ttx31可能编码碳水化合物结合结构域(carbohydrate binding module,CBM),但其结合特性未知.本实验以T. thermohydrosulfuricus为材料,通过菌落PCR扩增其Tts31序列,将获得的Ttx31克隆到表达载体pET22b(+)中得到重组质粒pEX31,序列分析表明,所得重组表达载体pEX31中的Ttx31序列正确.将重组载体pEX31转入大肠杆菌宿主表达菌Tuner中,IPTG诱导表达后纯化,采用SDS-PAGE电泳和非变性亲和凝胶电泳法,研究重组表达的目的蛋白TtX31与糖的相互作用.结果发现,TtX31能结合糊精,但不能结合可溶性淀粉、普鲁兰糖、支链淀粉等.实验结果可以认为TtX31是一种功能性的新碳水化合物结合结构域,可以结合糊精.     

青稞慢消化淀粉酶法制备技术研究

为了探索酶解处理对青稞慢消化淀粉含量的影响,以青稞淀粉为原料,分别以α-淀粉酶法,β-淀粉酶法,转葡糖苷酶协同α-淀粉酶处理法,转葡糖苷酶协同β-淀粉酶处理法等4种方法制备慢消化淀粉(SDS),通过单因素试验和正交试验确定最佳制备方案。结果表明,制备SDS的最佳方法为转葡糖苷酶协同β-淀粉酶处理法,具体处理条件为:β-淀粉酶的添加量0.032%,转葡糖苷酶的添加量0.192%,酶解时间8h,冷藏回生时间5d,淀粉乳浓度15%。此条件下制备的SDS含量为32.53%。本研究结果为青稞慢性消化淀粉产品的开发利用提供了科学依据和技术参考。     

外源酶和辣椒素对黑水虻处理厨余垃圾效率及幼虫生长的影响

从厨余垃圾成分和黑水虻幼虫的生活习性入手，通过添加外源酶以提升黑水虻处理厨余垃圾的效率和虫体质量。本研究尝试向厨余垃圾中添加淀粉酶和过氧化氢酶，以加速其中的淀粉分解，利于幼虫生长；并在2或5 d龄期幼虫培育阶段，向厨余垃圾中添加辣椒素刺激幼虫进食，以期提高厨余垃圾的处理速率和养殖虫体的粗蛋白质量等。研究表明，向厨余垃圾中添加质量分数0.1‰的α-淀粉酶和0.01‰的过氧化氢酶，并在5 d龄期幼虫培育阶段添加12.5‰的辣椒素可提高黑水虻处理厨余垃圾的效率、虫体鲜质量和干质量、粗蛋白和粗脂肪质量，较对照，黑水虻处理厨余垃圾速率提高了6.49%，其虫体鲜质量和干质量分别提高了29.70%和38.22%、粗蛋白质量提高了18.81%、粗脂肪质量提高了20.97%。微量外源酶（α-淀粉酶和过氧化氢酶）添加并在5 d龄期幼虫培育阶段加以辣椒素刺激，可显著提升黑水虻处理厨余垃圾的速率、虫体鲜质量和干质量以及粗蛋白和粗脂肪质量，研究结果可为厨余垃圾高效利用和高蛋白饵料基质的培育提供参考依据。     

Microbial degradation of profluralin

A selective culture technique combined with autoradiography was used to isolate microorganisms which could degrade profluralin [N-(cyclopropylmethyl)-α,α,α-trifluoro-2,6-dinitro-N-propyl-p-toluidine]. Of seven isolates obtained from a Cecil loamy sand, the three most active in degrading the herbicide were tentatively identified as Arthrobacter simplex, Cellulomonas ftavigenum and Microbacterium flavum. Degradation of profluralin was determined in pure and mixed culture in the dark and subsequently measured by TLC and scintillation methodology. M. ftavum produced three products which were identified as: 2-amino-α,α,α-trifluoro-6-nitro-N-propyl-p-toluidine; N-(cyclopropylmethyl)-α,α,α-trifluoro-2,6-dinitro-p-toluidine; and 2-amino-α,α,α-trifluoro-6-nitro-p-toluidine. The major product detected with all three organisms was 2-amino-α,α,α-trinuoro-6-nitro-p-toluidine. No degradation was detected in the absence of exogenous C and N.; co-metabolic processes appeared to be involved.     

Bacterial degradation of fungicide captan

The phthalimide fungicide captan has been widely used to control plant pathogenic fungi. A strain of Bacillus circulans utilized the fungicide captan as sole source of carbon and energy. The organism degraded captan by a pathway involving its initial hydrolysis to yield cis-1,2,3,6-tetrahydrophthalimide, a compound without fungicidal activity. The formation of this compound was confirmed by HPLC, IR, NMR, and mass spectral analysis. The results also revealed that cis-1,2,3,6-tetrahydrophthalimide was further degraded to o-phthalic acid by a protocatechuate pathway. These findings indicated that there was a complete mineralization of fungicide captan by B. circulans.     

Proxy global assessment of land degradation

Land degradation is always with us but its causes, extent and severity are contested. We define land degradation as a long-term decline in ecosystem function and productivity, which may be assessed using long-term, remotely sensed normalized difference vegetation index (NDVI) data. Deviation from the norm may serve as a proxy assessment of land degradation and improvement – if other factors that may be responsible are taken into account. These other factors include rainfall effects which may be assessed by rain-use efficiency, calculated from NDVI and rainfall. Results from the analysis of the 23-year Global Inventory Modeling and Mapping Studies (GIMMS) NDVI data indicate declining rain-use efficiency-adjusted NDVI on ca. 24% of the global land area with degrading areas mainly in Africa south of the equator, South-East Asia and south China, north-central Australia, the Pampas and swaths of the Siberian and north American taiga; 1.5 billion people live in these areas. The results are very different from previous assessments which compounded what is happening now with historical land degradation. Economic appraisal can be undertaken when land degradation is expressed in terms of net primary productivity and the resultant data allow statistical comparison with other variables to reveal possible drivers.     

磺胺嘧啶在水中的微生物降解研究

为了探明磺胺嘧啶在水中的环境行为,通过室内模拟降解实验分别研究了磺胺嘧啶在湖水和猪场废水中的好氧和厌氧微生物降解,考察了供氧方式和有机质含量对磺胺嘧啶微生物降解的影响。结果表明：磺胺嘧啶在猪场废水中厌氧微生物降解速率高于其好氧组,而磺胺嘧啶在湖水中厌氧微生物降解速率低于其好氧组。磺胺嘧啶在湖水和猪场废水中的好氧或厌氧微生物降解均较缓慢,这可能与其较强的抑菌性和微生物的营养状况有关。通过微生物培养还研究了好氧降解时磺胺嘧啶对湖水中微生物种群生长的影响,数据显示：磺胺嘧啶对湖水和猪场废水中细菌的生长具有一定的刺激作用,而对真菌和放线菌的生长影响不明显。     

Oxidative degradation of lipids during mashing

Although hardly any polyunsaturated fatty acids (PUFAs) are present in the endproduct, the ingredients used for the production of beer contain a high concentration of PUFAs, such as linolic and linolenic acid. These compounds are readily oxidized, resulting in the formation of lipid-derived products that reduce the taste and quality of beer enormously. During mashing relatively high amounts of PUFAs are exposed to atmospheric oxygen at a relatively high temperature. This makes mashing a critical step in the brewing process with regard to the formation of lipid-derived off-taste products. F1 phytoprostane (PPF1) changes in antioxidant capacity and monohydroxy fatty acids (OH-FAs) were used as markers for the detection of oxidative damage to fatty acids during mashing. The pattern of OH-FA formation indicates that enzymatic oxidation of PUFAs is more important than nonenzymatic oxidation during the mashing process. Nevertheless, substantial nonenzymatic radical formation is evident from the increase of specific OH-FAs and PPF1s. It was found that a low oxygen tension reduces oxidative damage and gives a high antioxidant capacity of the mashing mixture. This indicates that mashing should be done under low oxygen pressure.     

Thermal degradation studies of food melanoidins

Food melanoidins were isolated from bread crust, coffee, and tomato sauce and their composition was investigated by thermal degradation. Among the generated volatiles, important food flavor compounds were detected: in particular furans, carbonyl compounds, 1,3-dioxolanes, pyrroles, pyrazines, pyridines, thiophenes, and phenols. The results indicated that the isolated melanoidin fractions mainly consisted of compounds formed from carbohydrates and their degradation products. Besides proteins, other food constituents were incorporated in the melanoidin structure as well, such as lipid oxidation products in tomato melanoidins and phenolic compounds in coffee melanoidins. A comparison of the thermal generation of volatiles between these food-derived melanoidins and model melanoidins prepared from a single carbonyl compound and an amino acid showed that the degradation pattern of food melanoidins is quite different from that obtained from a glucose-glycine model system.     

Effect of gamma-irradiation on degradation of alginate

The aqueous solution of alginate was irradiated by 60Co gamma-rays in the dose range of 10-500 kGy. To assess the effect of irradiation on the degradation of alginate, the irradiation-induced changes in the viscosity, molecular weight, color, monomer composition, and sequence were measured. The molecular weight of raw alginate was reduced from 300000 to 25000 when irradiated at 100 kGy. The degradation rate decreased and the chain breaks per molecule increased with increasing irradiation dose. The viscosity of irradiated alginate solution reached a near minimum as low as at 10 kGy. No appreciable color changes were observed in the samples irradiated at up to 100 kGy, but intense browning occurred beyond 200 kGy. The 13C NMR spectra showed that homopolymeric blocks, MM and GG, increased and the M/G ratio decreased with irradiation. Considering both the level of degradation and the color change of alginate, the optimum irradiation dose was found to be 100 kGy.     

等离子体降解甲氰菊酯农药的效果

为探索高效快速降解甲氰菊酯农药的新方法,利用等离子体处理含有甲氰菊酯农药残留的山东寿光城阳青大白菜,研究了等离子体处理对甲氰菊酯农药残留降解效果的影响,采用响应面法对其影响因素进行了优化设计.结果表明:等离子体处理对大白菜中残留的甲氰菊酯降解有明显的效果,降解率均在70％以上,各参数影响顺序为:作用时间＞作用功率＞极距...