Simple Affinity Purification Method for Raw Starch-adsorbable and -digesting Amylases with a Raw Starch Column

A simple purification procedure for raw starch-adsorbable and -digesting amylases (RSAs) was devised. The method depended on an affinity column, which was prepared by mixing raw corn starch and Hyflo Super-Cel. RSAs were specifically adsorbed on the matrix, and eluted with a buffer containing 1% β-cyclodextrin. This column could be used to purify RSAs from Streptomyces thermo-cyaneoviolaceus and a recombinant strain of E. coli.     

Degradation of Raw Starch by a Wild Amylolytic Strain of Lactobacillus plantarum

Lactobacillus plantarum A6, isolated from fermented cassava, can break down cassava raw starch that has not been subjected to preliminary physicochemical treatment. When the pH was kept at 6, the microorganism cultured in a bioreactor excreted a high α-amylase activity (60 U/ml). Synthesis of the enzyme occurred during the stationary phase and resulted in full hydrolysis of the cassava starch granules. This gave 41 g of lactic acid from 45 g of raw starch after 3 days of fermentation. Enzymatic attack was evident under scanning electron microscopy in the rougher appearance of the surface of starch granules and in the presence of large cavities in some of them. In contrast, when the pH was not regulated, only a small amount of α-amylase activity was produced (2 U/ml) and no decrease in the starch content of the medium was observed. However, under scanning electron microscopy, some granules displayed a rougher surface, which might have been the result of weak enzymatic attack.     

一株降解生木薯淀粉的曲霉菌株的筛选、鉴定及其淀粉降解特性

本研究利用以生木薯淀粉为唯一碳源的筛选培养基,从腐烂木薯渣中分离筛选出一株可以降解生木薯淀粉的真菌菌株RSDF-7.根据RSDF-7形态和18S rDNA与28S rDNA之间的内转录间隔区(internal transcribed spacer,ITS)序列分析的结果,初步认定该菌株为曲霉属.菌株RSDF-7的粗酶液对多种不同的生淀粉底物均有水解效果;在以大米和玉米淀粉为底物时,其生淀粉分解活力比较高,分别为42%和40%.菌株RSDF-7的粗酶液具有良好的低pH稳定性,对生木薯淀粉的最适作用温度为50℃,最适作用pH为4.5.在30 min的吸附后,RSDF-7的粗酶液对生木薯淀粉的吸附力高达60%.使用HPLC对粗酶液的酶解产物进行检测,结果发现酶解产物中仅存在葡萄糖,表明菌株RSDF-7所产的生淀粉降解酶主要为糖化酶.扫描电镜观察结果发现,经RSDF-7粗酶液酶解后的生木薯粉颗粒破裂,形成空洞,说明RSDF-7粗酶液对生淀粉有较强的水解作用.可以预见,经纯化后的曲霉菌株RSDF-7生淀粉酶将来可以用于基于酶解的木薯淀粉转化.     

Floridean Starch

A survey was made of about 30 species of red algae from thePacific Coast to find the best starting-material for the isolation,in pure and native form, of the controversial substance, florideanstarch. Constantinea subulifera Setchell turned out to be theideal alga for this purpose. The isolated starches were subjectedto a number of physical, chemical, and enzymatic tests in orderto bring out possible differences from other starch-family substances,such as amylopectin and glycogen, isolated from higher plants.According to all criteria applied there is no real differencebetween the various compounds, except for the fact that florideanstarch gelatinizes only after very long boiling in water. End-groupdeterminations with the aid of periodate show that the florideanstarch molecule is a strongly branched structure somewhat comparableto glycogen.     

Improved Cassava Starch by Antisense Inhibition of Granule-bound Starch Synthase I

Cassava is a poor man's crop which is mainly grown as a subsistence crop in many developing countries. Its commercial use was first as animal feed (also known as tapioca), but has shifted since the late sixties to a source of native starch. The availability of native starches, which on the one hand do not require substantial chemical derivatisation and on the other hand have improved properties, would make cassava also for small farmers a potentially attractive cash crop. Since breeding is difficult in this polyploid, vegetatively propagated, crop a transgenic approach would be ideal to improve certain characteristics. We have created a cassava genotype producing amylose-free starch by genetic modification. The absence of amylose increased the clarity and stability of gels made with the transgenic starch, without requiring treatment with environment-unfriendly chemicals such as epoxides (propylene oxide, ethylene oxide) and acetic anhydride, which are normally used to improve stability. The amylose-free starch showed no changes in particle size distribution, chain length distribution or phosphorous content when compared to amylose-containing starch, but the granule melting temperature was increased by almost 2°C. Furthermore, the amylose-free cassava starch shows enhanced clarity and stability properties. These improved functionalities are desired in technical applications in paper and textile manufacturing, but also in the food industry for the production of sauces, dairy products and noodles.     

Spinnability of Starch Pastes

We have developed apparatus to assess spinnability that uses an impedance to prevent of electrolysis. The spinning distance and the stress confirmed by measurement of dial gauge.

A total of 10 starch pastes was prepared from corn, sago, katakuri, sweet potato, kuzu, edible canna, cassava, Indian lotus root, bracken, and potato, at 2% and 4% concentrations. The spinning distance of each starch paste showed a dependence on the concentration and the tensile velocity.

The strain at the point of maximum stress increased in all samples with increases in the tensile velocity. The spinning energy increased with increase in the tensile velocity, with a particularly large dependence on the tensile velocity of potato.

Cluster analysis was done using the spinning property values of 4% starch pastes as the parameters, which showed a division at a distance of 20, yielding four clusters, with each cluster showing a distinctive spinning pattern.     

Direct Production of Ethanol from Raw Corn Starch via Fermentation by Use of a Novel Surface-Engineered Yeast Strain Codisplaying Glucoamylase and α-Amylase

Direct and efficient production of ethanol by fermentation from raw corn starch was achieved by using the yeast Saccharomyces cerevisiae codisplaying Rhizopus oryzae glucoamylase and Streptococcus bovis α-amylase by using the C-terminal-half region of α-agglutinin and the flocculation functional domain of Flo1p as the respective anchor proteins. In 72-h fermentation, this strain produced 61.8 g of ethanol/liter, with 86.5% of theoretical yield from raw corn starch.     

Identification of Particles in Raw Materials

Raw materials need to be of a certain quality with respect to physical and chemical composition. They also need to have no contaminants in the form of particles because these could get into the product or indicate the raw materials are not pure enough to make a good quality product. When particles are found, it is important to identify their chemical and elemental composition to correct any process errors that can cause them and to have acceptable quality of the final product. Sources of materials can be the environment, process equipment and processing, and packaging. Microscope versions of Raman spectroscopy, laser-induced breakdown spectroscopy (LIBS), and IR spectroscopy are excellent tools for identifying particles in materials because they are fast and accurate techniques needing minimal sample preparation that can provide chemical composition as well as images that can be used for identification. The micro analysis capabilities allow for easy analysis of different portions of samples so that multiple components can be identified and sample preparation can be reduced or eliminated. The complementarity of the techniques provides the advantage of identifying various chemical components, as well as elemental and image analyses. The sources of materials were seen to be the environment, process equipment and processing, and packaging.     

小麦地方品种淀粉颗粒结合蛋白及淀粉含量差异研究

淀粉颗粒结合蛋白包含了多种淀粉生物合成的关键酶,对作物淀粉品质有重要影响。本研究利用1D-SDS-PAGE,分离了74份四川、西藏及云南毗邻地区小麦的淀粉颗粒结合蛋白,对突变材料进行了分子标记检测,对总淀粉和直链淀粉含量差异进行了比较。发现供试材料中,在分子量57～130 kDa区域共有9种不同的蛋白条带。其中,2个条带可能为新的淀粉颗粒结合蛋白;存在12份Wx-B1缺失的自然突变体和3份稀有的SGP-B1缺失突变体;筛选到直链淀粉含量超过30%的材料2份,直链淀粉含量为15%左右的材料10份。这些材料为小麦淀粉品质改良及淀粉生化合成机理研究提供了基础。     

淀粉含量不同的玉米自交系籽粒淀粉积累及其关键酶活性

以2个高淀粉和2个低淀粉玉米自交系为材料,分析了玉米籽粒淀粉的动态积累规律,同时对高低淀粉玉米籽粒灌浆过程中淀粉生物合成关键酶活性的动态变化及其与淀粉积累动态的相关性进行讨论分析。研究结果表明:灌浆过程中4个自交系淀粉含量变化趋势均呈sigmoid型曲线。灌浆过程中ADPG-PPase(腺苷二磷酸葡萄糖焦磷酸化酶)、SSS(可溶性淀粉合成酶)、GBSS(颗粒结合淀粉合成酶)活性均呈单峰曲线变化,峰值都出现在20～30DAP(授粉后天数)。2个高淀粉自交系的Q酶(淀粉分支酶)活性也呈单峰曲线变化,峰值也出现在20DAP,而2个低淀粉自交系的Q酶活性则呈双峰曲线变化,2个峰值分别出现在15～20DAP和30～35DAP。4个自交系籽粒淀粉的积累速率与各自交系ADPG-PPase、SSS和GBSS的活性变化呈极显著正相关。各自交系关键酶活性之间,ADPG-PPase、SSS和GBSS三者间活性变化呈极显著正相关,这3种酶活性变化与Q酶活性变化也呈不同程度的正相关。