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.     

基于氨基酸组成的黄酒酒龄电子舌鉴别

该研究采用电子舌结合化学计量学方法用于黄酒酒龄的快速鉴别。为确证黄酒样品酒龄,采用氨基酸分析仪分析了1年陈、3年陈和5年陈黄酒中20种氨基酸,并利用主成分分析（principal component analysis,PCA）对氨基酸数据进行了分析。采用电位型电子舌采集了不同酒龄黄酒样品的味觉指纹信息,并采用判别分析（discriminant analysis,DA）方法结合味觉指纹信息建立黄酒酒龄快速鉴别模型。采用偏最小二乘法（partial least squares regression,PLSR）建立电子舌响应信号与氨基酸含量之间的相关关系。氨基酸数据结合PCA分析表明所有样品均标注正确;电子舌结合DA所建黄酒酒龄鉴别模型可将3个年份预测集样品正确区分;异亮氨酸（Ile）、天门冬氨酸（Asp）、酪氨酸（Tyr）和缬氨酸（Val）与电子舌相关性高,模型的相对分析误差（Residual predictive deviation, RPD）高于2。研究表明电位型电子舌结合判别分析是黄酒龄鉴别的稳健方法。     

Chemical characteristics and potential source of fulvic acids leached from the plow layer of paddy soil

To understand better the chemical characteristics and source of dissolved organic matter (DOM) leached from the plow layer of rice paddies, a lysimeter study was conducted, which simulated submerged paddy topsoil during rice growth. The fulvic acid (FA) fraction in the percolation water from the lysimeter was collected by adsorption onto insoluble polyvinylpyrrolidone (PVP), and the temporal variations in its elemental composition, molecular size distribution, IR spectra, ¹³C CPMAS NMR spectra, and δ¹³C values were investigated. The proportion of the FA fraction to bulk DOM varied greatly, but the chemical characteristics of the FA fraction did not change appreciably during the experimental period. Thus, it is considered that the changes in the DOM composition in percolation water were mainly due to the differing contributions of the FA fraction. Further, to investigate the source of the FA fraction in the leachate, the chemical characteristics of the FA fraction in the leachate were compared with those extracted from the plow layer soil. A sequential extraction of the FA fraction was conducted using a sequence of water, 0.25 M Na₂SO₄, 0.1 M Na₄P₂O₇ (pH 7.0), 0.1 M Na₄P₂O₇ (pH 10.5), and NaBH₄+0.1 M Na₄P₂O₇ (pH 10.5). It was found that the water- and 0.25 M Na₂SO₄-extractable fractions, which were most mobile, were not the only source of the FA fraction in the leachate. The small molecular size sub-fraction of the NaBH₄+0.1 M Na₄P₂O₇ (pH 10.5)-extractable FAs, most of which are probably bound mainly to iron oxides, are considered to be another source of the FA fraction leached from the plow layer of paddy fields.     

表层土壤中菲的紫外光降解研究

多环芳烃化合物（PAHs）由于致癌、致畸和致突变而受到广泛关注。本实验以多环芳烃菲（Phe）为目标污染物,研究了温度、腐植酸和紫外辐射强度对Phe光降解的影响并对不同因素对降解动力学的影响作了研究。结果表明,Phe的降解在20 ℃到30 ℃范围内,随着温度的升高,光降解率增加;腐植酸在Phe污染土壤的光降解中起敏化作用,可显著促进光解,腐植酸浓度为5 mg·kg^-1足以达到敏化的效果;Phe光降解速率常数随辐射强度的降低而降低,呈正相关,光解的半衰期随着辐射强度的降低而增加,呈负相关。     

不同硼营养下甘蓝型油菜体内游离氨基酸成分变化分析

本文研究了不同供硼水平（缺硼、正常供硼、硼过量）对油菜幼苗叶片及花药内游离氨基酸成分的影响。实验结果表明，缺硼和硼过量都会造成植株生长异常，幼苗干物重显著下降，植株幼苗叶片与花药内蛋白水解酶的活性升高，而游离氨基酸总量增加，同时，氨基酸的组分也发生了明显的变化。不适宜供棚（缺硼、硼过量）使幼苗叶片内辅氨酸含量明显升高，与之相伴随的是一些氨基酸含量的下降，而花药内脯氨酸含量有所减少。     

双低油菜籽粕营养成分及饲喂育成鸡研究

本文分析了双低油菜籽粕的营养及抗营养成分，并以不同的日粮比例对０～２０周龄蛋鸡进行饲养试验。结果表明，双低油菜籽粕与普通菜籽粕的粗蛋白、氨基酸含量相似，硫甙及其水解产物ＯＺＴ，ＩＴＣ等抗营养成分仅为１／１０左右。在日粮中添加１０％，１５％双低油菜籽粕，对蛋鸡生长发育、甲状腺及血清Ｔ３，Ｙ４，ＴＳＨ，ＴＧ，Ｔｍ水平，肝及血浆ＣＨＥ，ＡＫＰ，ＧＯＴ，ＧＰＴ的活性和肾脏等均无异常影响。日粮中添加２０％双低油菜籽粕，鸡生长明显受到抑制，甲状腺肿大，血清Ｔｃ上升，Ｔ３下降，肝出血明显，血浆中ＧＯＴ，ＧＰＴ明显升高。故认为，双低油菜籽粕是１种氨基酸组成较均衡、毒素含量较低的蛋白质饲料，在０～２０周龄蛋鸡日粮中加到１５％时均安全。     

Atmospheric carbon dioxide enrichment reduces carbohydrate and nitrogen reserves in overwintering Picea mariana

Abstract

Elevated levels of atmospheric carbon dioxide (CO₂) can directly affect the cold hardening process in evergreens through their effect on the accumulation of carbon and nitrogen reserves. This study investigated the biochemical responses of black spruce [Picea mariana (Mill.) B.S.P.] seedlings to CO₂ enrichment during growth, cold hardening and dehardening. Seedlings were grown under 350 (ambient) or 710 (elevated) ppm of CO₂ for 12 months in eight mini-greenhouses. Photoperiod and temperature were gradually lowered in autumn to induce cold hardening, and the conditions were reversed in spring to promote dehardening. At regular intervals, cold tolerance was assessed and sugars, starch and amino acid concentrations were measured. The freezing tolerance differed between the two treatments only in early autumn, with seedlings growing under high CO₂ being more tolerant. The northern ecotype was more cold tolerant with concomitant higher concentrations of sucrose, fructose, pinitol, glucose and total soluble sugars. The concentration of soluble sugars increased in needles and roots of black spruce along with cold hardening, and the concentrations of the cryoprotective sugars sucrose and raffinose were lower under elevated CO₂. Amino acid concentrations were also lower under elevated than under ambient CO_2. The lower level of reserve did not translate into a lower level of freezing tolerance.     

微波对毛酸浆籽油提取效率的影响及其脂肪酸分析

以毛酸浆种子为原料,研究索氏微波提取法对毛酸浆籽油提取效率的影响,并以甲酯化脂肪酸为标准品,采用气相色谱法对毛酸浆籽油脂肪酸成分进行分析,通过与标准品比对出峰时间确定试样脂肪酸成分,使用面积归一法确定试样中各脂肪酸含量。结果表明,与普通热风干燥相比,微波干燥法能够提高毛酸浆籽油的提取效率;利用气相色谱法对毛酸浆籽油进行脂肪酸分析,共检测出8种脂肪酸,分别为:肉豆蔻酸1.3%,棕榈酸10.3%,硬脂酸5.3%,油酸14.3%,亚油酸62.7%,亚麻酸4.0%,花生酸1.1%,二十二烷酸1.0%。其中不饱和脂肪酸相对含量占81.0%,说明毛酸浆籽油具有较高的营养价值,开发前景广阔。     

115份贵州茶树资源氨基酸和茶氨酸分析与特异资源筛选

为筛选高氨基酸和高茶氨酸茶树资源,探明贵州茶树资源氨基酸和茶氨酸特性,本研究分别利用分光光度法和高效液相色谱法测定115份贵州地方茶树资源春季一芽二叶生化样的氨基酸总量和茶氨酸含量。结果表明：115份茶树资源的氨基酸总量在0.95%～7.96%,平均值为3.40%;茶氨酸含量在0～3.80%,平均值为1.58%;茶氨酸占比在0～67.66%,且集中分布于40%～<60%。在这115份茶树资源中,发现特异高氨基酸(氨基酸总量≥5%)茶树资源11份、特异高茶氨酸(茶氨酸含量≥3%)茶树资源3份。不同地方的茶树资源,其氨基酸总量和茶氨酸含量存在较大差异,其中,贵定、黎平和石阡的茶树资源表现出较高的氨基酸总量和茶氨酸含量,而三都和普安的茶树资源氨基酸总量和茶氨酸含量显著(P<0.05)低于其他地方。来自三都的突肋茶(Camellia costata)资源表现出更原始的生化性状,均未检测出茶氨酸。聚类分析结果显示,115份茶树资源可分为5类,其中,石苔14号以远高于其他资源的氨基酸总量和茶氨酸含量单独分为一类。     

盐水鸭加工过程中滋味成分变化研究

该文采用高效液相色谱和氨基酸自动分析仪研究了盐水鸭加工过程中的滋味成分变化.盐水鸭加工过程中大部分小肽含量具有减少的趋势.在煮制前的加工中,游离氨基酸含量增加而风味核苷酸含量减少,煮制过程中两者的含量均显著减少.干腌后鸭肉含盐量最高,但经其后工序加工后含量降为适宜食用的3%左右.重要的滋味成分盐水鸭含量均高于对照鸭肉.风味核苷酸和鲜味氨基酸对盐水鸭的滋味具有重要贡献.盐水鸭加工过程中复卤工艺对鸭肉滋味成分作用显著,是构成盐水鸭美味的原因之一.