短小芽孢杆菌HB-3的分离鉴定及其淀粉酶的纯化特性研究

从湖北省襄阳市三九酿酒厂下水道污泥分离筛选得到产淀粉酶菌株HB-3,根据菌株生长形态和生理生化特征分析,初步鉴定为短小芽孢杆菌。短小芽孢杆菌HB-3在淀粉培养基上培养72 h后,淀粉酶活力达到120.0 U/mL。然后使用DEAE-52和CM-52阴阳离子交换层析柱对短小芽孢杆菌HB-3产生的淀粉酶进行了部分纯化。结果表明,短小芽孢杆菌HB-3中淀粉酶经过部分纯化后,纯化倍数为5.9,酶活力回收率为90.9%。纯化后的淀粉酶使用SDS-PAGE凝胶电泳研究,以标准蛋白为对照,计算出短小芽孢杆菌HB-3中淀粉酶分子量为56.5 kDa。同时对短小芽孢杆菌HB-3的淀粉酶进行了纯化特性研究,结果表明,该酶的酶促反应最适温度为50℃,40～60℃热稳定性良好;该酶的酶促反应最适pH为5.0,pH 4.0～6.0酸碱稳定性良好,该酶对底物淀粉的反应动力学米氏常数K_m为0.062 mmol/L,最大反应速率V_(max)为3.019 mmol/(L·min)。     

胰α-淀粉酶基因5''端调控序列与鱼类食性的关系

为了研究不同鱼类胰α-淀粉酶基因5'端调控序列转录因子差异与鱼类食性分化之间的关系。通过PCR克隆测序和查询NCBI数据库,获得32种鱼类胰α淀粉酶基因5'端824 bp序列,并对鱼类胰α淀粉酶基因5'端序列进行转录因子和系统发育分析。按不同的营养类型将鱼类分为杂食性、植食性和肉食性,通过百分比相似性分析不同食性鱼类的胰α淀粉酶基因转录因子的组成差异以及转录因子与鱼类食性的关系。结果显示:不同食性鱼类的胰α-淀粉酶基因5'端序列存在转录因子种类差异,植食性-肉食性鱼类差异主要体现在E47、C/EBPalpha、NF-Y和Pax-2,植食性-杂食性差异主要体现在deltaEF1、MyoD、NF-Y、AREB6和Pax-2,杂食性-肉食性差异主要体现在GATA-1、SRY、MyoD、HFH-8、AREB6、Pax-2、STAT5A和AP-1。系统发育结果与传统形态分类学大体相符,相同食性的鱼类并没有聚为一类。鱼类胰α-淀粉酶基因5'端调控序列中与食性分化相关的转录因子有E47、C/EBPalpha、NF-Y、Pax-2、deltaEF1、MyoD、AREB6、GATA-1、SRY、HFH-8、STAT5A和AP-1;胰α-淀粉酶基因5'端序列的转录因子与鱼类食性分化具有一定的关系。     

Isolation and Identification of Producing High-yielding Amylase Clostridium butyricum from Chicken

In order to obtain a producing amylase Clostridium butyricum,the chicken small intestinal mucous was collected to isolate and screen Clostridium butyricum.First of all,the samples were heated at 80℃ for 10 min for removing most non-spore bacteria,then the samples were inoculated on TSN medium.The suspected Clostridium butyricum strains were screened by colony morphology and microscopic morphology observation.The producing acid ability and amylase activity of the isolates were successively tested,so the C.B1 strain,both producing acid and producing high-yielding amylase,was selected to be identified by physiological and biochemical test and 16S rDNA sequence analysis.The results showed that C.B1 strain had the characteristics of gram-positive anaerobe,rod-shaped and forming a circular or oval spore,16S rDNA sequence was 1450 bp.Homology comparison analysis indicated that it was most closest to Clostridium butyricum with 99% homology.So C.B1 strain was identified as Clostridium butyricum,this study would lay the foundation for developing new microecological preparations.     

Ontogenic development of digestive enzymes in bigfin reef squid (Sepioteuthis lessoniana)

Bigfin reef squid (Sepioteuthis lessoniana) is an economically important exported aquatic animal in Thailand, but little is known about its husbandry, which needs to be clarified. In this study, the ontogenic development of the main digestive enzymes (pepsin, trypsin, chymotrypsin, lipase and amylase) was investigated within 30 days after hatching (DAH). Fertilized eggs of the wild cuttlefish were hatched and then distributed into four replicate aquaria assigned to varying collection times (0, 5, 10, 15, 20, 25 and 30 DAH). The development of pepsin suggests acidic digestion functionality at 5 DAH, and its specific activity was maintained over the studied period. Specific activities of the serine proteases, trypsin and chymotrypsin, fluctuated significantly with similar pattern, and appear to be the key enzymes contributing ontogenic changes in protein catabolism. The specific activity of lipase was relatively high at 0 DAH, dramatically increased by 5 DAH, and then appeared constant until the end of observations, suggesting important role in utilizing the lipid from yolk at hatching, as well as probably in sparing protein utilization. Dramatic increase in amylase specific activity occurred by 10 DAH after which the activity was constant until 30 DAH, suggesting the utilization of carbohydrate as energy evolves comparatively late. Findings from the current study could be used to manage nutritional protocols according to the digestive enzyme patterns, as well as to develop artificial diets for rearing S. lessoniana.     

Assessment of prebiotic potentials in selected leaf meals of high dietary fibre on growth performance,body composition,nutrient utilization and amylase activities of a tropical commercial carp fingerlings

The performance of selected leaf meals of high dietary fibre in the feed of a tropical commercial carp, hybrid lemon fin barb (Barbonymus gonionotus ♀ × Hypsibarbus wetmorei ♂) was evaluated in a 56‐day feeding trial. The tropical carp juveniles (9.43 ± 0.05 g) were randomly stocked in 60‐L aquaria at 15 fish per aquarium. Five isonitrogenous and isocaloric diets (30% crude protein, 17 kJ/g gross energy) containing no leaf meal and 10% napier grass (Pennisetum purpureum), alfalfa (Medicago sativa), water spinach (Ipomea aquatica) and Gliricidia sepium leaf meals were formulated and tested. Three aquaria were randomly assigned to each experimental diet. The results showed that fish‐fed diets containing leaf meals showed superior growth performance and body composition which was better than those fed control diet. The leaf meal‐treated groups also had higher values of protein, lipid, energy retention and production of digestive enzymes amylase. Conclusively, the results indicated that leaf meal fibre provided better performance showing the inherent prebiotic effect of the utilization of these leaf meal in hybrid lemon fin barb .     

山体库贮藏马铃薯淀粉、还原糖含量及淀粉酶活性的变化

为探讨甘肃省民乐县山体库贮藏对马铃薯品质的影响。对9个马铃薯品种不同贮藏时期淀粉、还原糖含量以及淀粉酶活性进行分析测定。结果表明：随着贮藏期的延长，大多数品种的淀粉含量表现为升→降→升→降，还原糖含量表现为升→降，淀粉酶活性表现为升→降→升→降；2月份是一个临界贮藏时期，淀粉含量降低到最低（12.68%~22.47%），还原糖含量达到最高点（0.43%~2.41%），淀粉酶活性降低到最低点（0.43~0.87 mg/(g? min)）。山体库有利于马铃薯的贮藏；‘青薯6号’和‘同薯23号’适合做菜用型品种，‘大西洋’、‘夏波蒂’和‘LK-99’适合做生产加工型品种，为了保证品质，最佳的加工时间在次年2月之前完成；淀粉酶活性和淀粉含量之间存在负相关，与还原糖之间存在正相关，淀粉含量和还原糖含量之间存在负相关，淀粉含量和还原糖含量之间处于动态平衡状态。     

A rapid,automated method for measuring α-amylase in pre-harvest sprouted (sprout damaged) wheat

The quality of wheat for baking is critically dependent on the level of α-amylase (1,4-α-D-glucan glucanohydrolase, EC 3.2.1.1), which can be present as “late maturity α-amylase” (LMA), or due to pre-harvest sprouting due to high rainfall and humidity at the time of harvesting. The most commonly used method to measure α-amylase in wheat grain is the Hagberg Falling Number method, but values are also influenced by rheological properties of starch in the grain. In this study we describe a simple, rapid, automated method (Amylase SD) for measurement of α-amylase in pre-harvest sprouted (sprout damaged) wheat grain. The method (Amylase SD) measures the release of p-nitrophenol from 4,6-O-ethylidene-α-4-nitrophenyl-maltoheptaoside by α-amylase in the presence of α-glucosidase. The absorbance of p-nitrophenolate measured at 405 nm in a ChemWell^®-T auto-analyser is directly related to the level of α-amylase activity present in the milled wheat grain extract. The Amylase SD method generated <6%CV and correlation to the Falling Number method was represented by an inflection point at ∼160 s. The precision, sensitivity and speed of this method provides an ideal alternative to the Falling Number method for measurement of α-amylase (sprout damage) in wheat grain in wheat breeding programmes or at grain receival points.     

Effect of two wheat genotypes and Swedish environment on falling number, amylase activities, and protein concentration and composition

Variation in falling number, amylase activities, protein concentration and composition were investigated in two wheat cultivars grown in Sweden over two seasons, in four locations, with four N fertilizer rates, with and without fungicide treatment. The results showed that;