转Bar、Bt基因双抗大米外源基因在食品加工过程中稳定性的研究

旨在了解转Bar、Bt基因双抗大米外源基因在酒酿和米果加工过程中的稳定性,以转Bar、Bt基因双抗大米为原料,进行了6种不同方式的加工,并针对其加工品中外源基因(包括CaMV35S启动子、Ubiquitin启动子、NOS终止子、Bar基因和Bt基因)进行不同大小片段的PCR扩增。结果表明,实验中长度小于200bp的片段均具有较高的GC含量(NOS终止子除外)和良好的稳定性。      

利用PCR方法检测水稻及其加工产品中转基因水稻

市场上的转基因产品以及深加工产品越来越多,其安全性引起全世界的极大争论,很多国家的检验机构相继开展了转基因产品的检测工作。以转基因水稻为材料,以聚合酶链式反应(PCR)方法为基础,选择适用于转基因食品安全性检验的核酸检测技术,针对转入苏云金杆菌(Bacillus thuringiensis,简写为Bt)基因水稻的Cry1Ac片段进行PCR检测,建立适合转基因水稻的检测方法。该方法简便快速、检测结果与标准及其他文献资料相符。     

铁蛋白转基因水稻与亲本的稻米营养成分的比较研究

对3份转Fer基因水稻与亲本的稻米营养成分进行了比较与分析,发现其中2份转Fer基因水稻(F2-5和F10-4)铁含量为野生型亲本秀水11的2.21和2.18倍,由此表明,铁蛋白基因表达与稻米铁含量变化有一定相关性。进一步鉴定表明,转基因稻米其它矿质元素以及粗蛋白、粗脂肪、淀粉、粗纤维、总灰分、氨基酸、维生素等关键性营养成分与亲本差异不显著,转基因未改变原亲本的其他品质性状。并就转Fer基因稻米的食品安全性以及转入的外源基因等问题进行了讨论。     

An investigation of two xylan‐degrading enzymes and a novel xylanase inhibitor in malted barley

Two enzymes with activity on xylan and xylan‐based substrates were isolated from malted barley. One appears to be relatively anionic and the other more cationic. Both are endo‐acting enzymes. The first of these was studied in more detail and shown to be of molecular weight ca. 59,000 and capable of acting through the pH range 5–7.5. It is relatively heat tolerant, with 30% of its activity surviving for 30 min at 70 °C. A low‐molecular‐weight (ca. 1300) inhibitor of xylanase that developed during germination was investigated. Its potency was not lowered by treatment with proteolytic enzymes, but treatment with polyvinylpolypyrrolidone did reduce its impact. Ultraviolet spectrophotometry and Fourier transform infrared spectroscopy, as well as staining with Folin–Ciocalteu reagent, indicated the presence of phenolic species, while mass spectroscopy studies suggested the presence of arabinose and possibly poly‐hexose. The presence of material that stained with aniline phthalate would be consistent with the presence of reducing sugar in the inhibitor. Copyright © 2013 The Institute of Brewing & Distilling     

Toxicological evaluation of transgenic rice flour with an Escherichia coli phytase gene appA by sub‐chronic feeding study in Wistar rats

BACKGROUND: Phytase is increasingly used as an ingredient in swine and poultry feeds to improve the bioavailability of phytate‐bound phosphorus and reduce the supplement of elemental phosphorus in feeds. Recently, a transgenic rise (TGR) with phytase appA gene from E. coli has been developed in Taiwan. To assure the food safety of this TGR, we performed a sub‐chronic whole‐food feeding study with Wistar rats. RESULTS: Weaned feeding male and female rats were divided into TGR and non‐TGR (NTGR) groups (20 rats per group), and fed diets containing 76.8% rice flour of TGR and NTGR, respectively. After 93 days feeding, although some hematological findings and blood chemistry values differed significantly between the TGR and NTGR groups, all values were still within the normal range for rats of this age and sex. No adverse effects of TGR were observed in terms of animal behavior, weight gain, or feed utilization rate. Necropsy at the end of the experiment indicated that neither pathological lesions nor histopathological abnormalities were present in organs such as liver, kidney, intestines and testes of rats in TGR and NTGR groups. In addition, no significant differences (P > 0.05) were observed in relative organ weights, hemograms and blood indices of rats between the two groups. CONCLUSION: These results demonstrated that in a 93‐day feeding study with Wistar rats a diet containing 76.8% of TGR flour having a phytase activity of about 1500 U kg^?1 body weight per day has no adverse effects. Copyright © 2007 Society of Chemical Industry     

外源酶改善糙米食味品质的研究

实验通过用纤维素酶、复合果胶酶来部分降解糙米中的粗纤维和果胶物质,改善糙米的食用品质。通过测定纤维素等物质的变化及糙米饭的感官,确定纤维素酶的作用条件为:反应温度45~50℃,加酶量0.11mg/mL,pH值4.8~5.2,作用时间4h;复合果胶酶的作用条件为:反应温度45~55℃,pH值5.0~5.6,加酶量0.6mg/mL,酶作用时间2h以上。实验发现经纤维素酶和复合果胶酶处理后的糙米食用品质较处理前有明显改善,其中纤维素酶作用好于复合果胶酶,混合酶作用效果最好。     

植物生长调节剂对烤烟内源激素、烟碱、多酚含量的影响

植物生长调节剂对烤烟内源激素以及烟碱、多酚含量的影响结果表明,植物生长调节剂GA、IAA、2,4-D不同程度地提高了内源GA、IAA含量,降低了内源ABA含量,其中外源GA、高浓度2,4-D处理效果较为明显;植物生长调节剂ABA较强地提高了内源ABA的含量,而降低了内源GA、IAA的含量,喷施高浓度ABA效果明显,在打顶后30d,内源ABA提高140%,而内源GA、内源IAA分别降低了61.9%、85.0%,内源GA在打顶后20d含量达到高峰,内源IAA、ABA在打顶后30d达到高峰。      

产木聚糖酶毕赤酵母工程菌株构建及表达条件优化研究

将黑曲霉XZ-3S木聚糖酶Xyn43A成熟肽基因插入表达载体p PIC9K,重组质粒SalⅠ线性化后分别电击转化2种毕赤酵母GS115和KM71,转化液经MD平板、G418浓度梯度平板和摇瓶复筛,获得两株重组菌GS115/Xyn43A（Mut⁺）和KM71/Xyn43A（Mut^s）。其中GS115/Xyn43A菌株最优表达条件为:甲醇浓度2.0%,接种时间24 h,诱导时间108 h,诱导温度30℃、诱导培养基初始p H6.3;KM71/Xyn43A菌株最优表达条件为:甲醇浓度1.75%,接种时间26 h,诱导时间132 h,诱导温度30℃、诱导培养基初始p H6.5。在最优表达条件下两重组菌GS115/Xyn43A和KM71/Xyn43A比酶活力分别可达139.36、143.29 U/mg。      

转Bt基因大米暴露对亲代雌性大鼠免疫系统影响的研究

目的 研究转Bt基因大米(TT51)对亲代雌性大鼠免疫系统的影响.方法 试验分为转Bt基因大米组(TT51组)、亲本明恢63大米组(明恢63组)和市售大米组,各组大米按60％的比例掺入饲料进行全食物喂养.Wistar雌、雄大鼠喂养10周后进行交配繁殖,对亲代雌性大鼠于断乳后进行免疫毒性评价.指标包括血常规、全血淋巴细胞分型、脾细胞的NK活性检测、刀豆蛋白A诱导淋巴细胞转化试验、抗体生成细胞检测以及淋巴结的淋巴细胞分型.结果 脾淋巴细胞分型中,TT51组的Th细胞、T细胞比例与市售大米组比较差异有统计学意义(P＜0.01),明恢63组的B细胞、T细胞比例与市售大米组比较差异有统计学意义(P＜0.01);颈部淋巴结淋巴细胞分型中,TT51组的Th细胞比例与市售大米组比较差异有统计学意义(P＜0.01);各组其他功能性试验结果之间差异无统计学意义.结论 转Bt基因大米(TT51)对亲代雌性大鼠免疫系统功能未见不良影响.     

米胚芽中胰脂肪酶抑制剂的分离纯化

由米胚芽中提取的胰脂肪酶抑制剂经鉴定主要为蛋白质.通过硫酸铵分级沉淀(40%～80%)、凝胶层析(Sephadex G-75)对米胚芽水溶物进行分离纯化.所得的洗脱峰I和洗脱峰Ⅱ的组分进行高效液相色谱分析,测定了胰脂肪酶活性抑制率较高的组分的分子量,分别为125KD和21KD.     

内源性物质对不同品种大米淀粉消化性的影响

近年来培育出了低高血糖生成指数（GI）大米品种,但是其产生低GI作用机理并不明确,内源性非淀粉物质作为大米关键成分,对不同GI大米淀粉消化率的影响值得探讨。本研究以典型的两种不同GI大米作为研究对象,探究了去除内源性非淀粉物质对大米淀粉体外消化性、理化性质和微观结构的影响。结果表明,对于低GI品种,蛋白质和膳食纤维的去除显著促进了淀粉消化,使最终消化率提升了7.37%~13.28%,但高GI品种的内源性非淀粉物质对于淀粉消化性的影响并不显著。在理化性质上,蛋白质和膳食纤维的去除将淀粉的膨胀力显著提升至17.83±0.30g/g~13.44±0.27g/g,显著改变了米粉的糊化特性和热力学特性,从而使其更容易糊化和消化。在微观结构上,去除蛋白质和膳食纤维使淀粉颗粒结构减小了3~8 μm,蛋白质、膳食纤维和淀粉的相互作用提高了米粉的有序度和聚集结构。     

应用响应面法优化黑曲霉固态发酵稻谷壳产木聚糖酶条件

对黑曲霉产木聚糖酶发酵工艺进行优化研究。在单因素实验的基础上,采用Box-Benhnken实验设计,对黑曲霉发酵稻谷壳产木聚糖酶的最佳碳源及其添加量、培养时间、水的添加量进行优化,建立了木聚糖酶产量随蔗糖添加量、培养时间和水的添加量变化的多项二次回归方程,并得到最佳的发酵工艺条件是:培养时间为78h,水的添加量为57%,蔗糖添加量为1.3g时,木聚糖酶的酶活力可达221.5U/g,比优化前提高了1.76倍,预测模型可靠,可利用该模型对黑曲霉产木聚糖酶的最优发酵条件进行理论预测。      

Loop结构引入半胱氨酸对木聚糖酶XynASP热稳定性的影响

为了探究Loop结构引入半胱氨酸对GH11家族木聚糖酶热稳定性的影响,以溶糖曲霉（Aspergillus saccharolyticus）JOP 1030-1木聚糖酶XynASP Loop结构为研究对象,通过定点突变技术,将第95位点天冬酰胺（Asn）突变成半胱氨酸（Cys）,获得突变体XynN95C,并在大肠杆菌（Escherichia coli）BL21（DE3）中诱导表达。酶学性质分析结果表明,突变体XynN95C最适温度为50 ℃,酶活性半衰期t1/240 ℃为38 min,相比野生型XynASP分别提高了5 ℃、18 min,最适pH从6.0降至5.0。另外,XynN95C的金属离子耐受性较强,经Fe3+处理1 h,相对酶活性为93.9%,较野生型（65.9%）明显提高。由此得出,在Loop结构引入半胱氨酸可有效提高木聚糖酶的热稳定性,这为GH11家族木聚糖酶的热稳定性改造又提供一新思路。     

固态发酵啤酒糟产木聚糖酶的菌种筛选及其产酶条件研究

从40株保藏菌种中筛选得到1株能发酵啤酒糟高产木聚糖酶的菌株黑曲霉AN27—2—1。通过单因素实验和正交实验对黑曲霉AN27—2—1固态发酵啤酒糟产木聚糖酶的条件进行优化，结果表明：在培养基组分为啤酒糟：玉米芯：麸皮=6：2：2，NH4NO3 2％，吐温800.1％，加水比为1：2，接种量为3．5％，培养温度为30℃，培养周期为2d-3d，产酶活达310．54U／g。     

黑曲霉木聚糖酶（xynZF-2）基因克隆、表达及酶学性质分析

利用RT-PCR技术从黑曲霉（Aspergillusniger）XZ-3S中成功克隆出木聚糖酶基因（xynZF-2）的cDNA序列（Genbank登录号为：JQ700382）。该基因核苷酸序列阅读框全长678bp,编码225个氨基酸,其中成熟肽为107个氨基酸。以重组质粒pUCm-T—xynZF-2为模板,扩增得到编码xynZF－2成熟肽的cDNA片段（624bp）。将其与表达载体pET－28a连接,构建重组表达载体pET－28a—xynZF－2,并转化大肠杆菌（Escherichiacoh）BL21（DE3）,获得重组工程菌BL21／xynZF-2。经IPTG诱导表达,木聚糖酶的的比酶活最高可达42．33U／mg。重组表达的木聚糖酶最适反应温度为40℃,最适反应pH为5．0,在碱性条件下具有良好的稳定性。Fe^3＋对酶的激活作用最为明显。     

盐析和双水相萃取法纯化细菌木聚糖酶的比较

通过对比硫酸铵盐析和双水相萃取两种木聚糖酶初级纯化方法的纯化效率,确定更加适于Paenibacillus sp.B1709木聚糖酶初级纯化的方法。结果显示,双水相萃取较盐析具有更好的木聚糖酶纯化回收效果,并对其主要实验参数进行优化,得出双水相萃取最佳条件为:聚乙二醇(PEG)4000 Da,浓度22%,最佳成相盐(NH₄)₂SO₄,最适浓度18%,最适pH6.0,NaCl 2%。在此条件下,可实现分配系数9.90,酶活力回收率为90.84%,纯化倍数6.13。实验结果显示双水相萃取相较于盐析在大规模纯化尤其针对复杂纯化目标时具备一定技术的优势。     

粳米与籼米组合对婴幼儿米粉食用品质的影响

研究了粳米和籼米不同搭配组合的基本理化指标、蒸煮特性、糊化特性和质构特性.结果表明,直链淀粉含量对食用品质具有重要影响.蒸煮特性上,直链淀粉含量越低,吸水率及膨胀体积越小,米汤pH、碘显色值和干物质含量越高,蒸煮及冲调性越好;糊化特性上,直链淀粉含量越低,峰值黏度和崩解值越高,回复值及衰减值越低,口感越好;质构特性上,...     

Effects of gelatinisation level, gum and transglutaminase on the quality characteristics of rice noodle

The aim of the present study was to investigate the effects of gelatinisation level, gum (locust bean gum, xanthan gum, 3%) and/or transglutaminase (TG, 0.5%) on quality characteristics of rice noodle. In order to improve the dough forming ability, rice flour was gelatinised at levels of 15%, 20%, 25% and 30%. Noodle samples were evaluated in terms of cooking loss, total organic matter (TOM), water absorption, swelling volume, maximum force, colour, sensory properties, pasting properties. Noodle sample with a gelatinisation level of 25% had better cooking and sensory properties. Gum and/or TG were added to this noodle formula. The noodle samples including xanthan gum had better cooking and sensory properties. TG caused a significant decrease in TOM. The samples including locust bean gum had significantly higher maximum force values. Xanthan gum caused decreases in some Rapid ViscoAnalyzer viscosity values of the noodle samples, while locust bean gum caused increases.     

Comparative physicochemical properties and structure of rice containing the sck+cryIAc genes and its nontransgenic counterpart

ABSTRACT:  The physicochemical properties and structure of an insect-resistant rice, Liangyou Kefeng Nr. 6 (IRR), containing the sck and cryIAc genes were compared with those of its nontransgenic counterpart designated as Liangyou 2186 (control), considering their key role in determining commercial value. Basically the appearance of IRR was not affected in terms of size and shape after foreign gene transformation but improved with lower chalkiness. The milling yield of IRR with lower chalkiness was higher measured by head rice yield compared with its parental control. The differences in appearance and milling quality were confirmed by the structure of raw rice grain by scanning electron microscopy (SEM). Slight differences were observed in pasting properties and textural quality determined by rapid viscosity analyzer and texture analyzer which was in agreement with the result of the structure of cooked rice grain by SEM. The above differences might be as a result of a positional effect of T-DNA insertion. On the whole, the appearance, milling quality, and eating quality of IRR were not adversely affected by transgenes, which will facilitate its acceptance by the consumer after commercialization.     

Effects of rice properties and emulsifiers on the quality of rice pasta

Two commercial rice flours (GTIndica and GTJaponica, with amylose contents of 306.8 and 185.2 g kg^?1 respectively) and their blended flours were used to make rice pasta. The effects of two emulsifiers (distilled glyceryl monostearate (GMS) and a commercial emulsifier (KM3000)) on the pasting and thermal properties of dried rice pasta were investigated using a Rapid Visco Analyser (RVA) and a differential scanning calorimeter (DSC). The rice pasta made from high‐amylose rice flour had better extrusion properties, better texture, whiter colour, less cooking loss and better eating quality than that made from low‐amylose rice flour. The maximum amount of low‐amylose rice flour that could be blended in for making an acceptable quality of rice pasta was 500 g kg^?1. A soft texture, low adhesiveness and low gumminess of cooked rice pasta were obtained by adding 10 g kg^?1 GMS. SEM investigations showed that use of an emulsifier restricted the swelling of starch granules, especially for the pasta made from high‐amylose rice flour. © 2001 Society of Chemical Industry