影响类弹性蛋白多肽自组装成微球的因素及其作用机制

影响类弹性蛋白多肽(ELPs)自组装成微球的因素较多,目前尚缺乏系统研究。以类弹性蛋白多肽[KV8F]n为对象,利用动态光散射仪测定了不同条件下其自组装成微球的粒径。结果表明:随着分子量的增加ELPs形成的微球粒径也随之增大,粒径的均一度减小;当盐浓度低于0.4 mol/L时,盐浓度的增加,微球粒径相应增加,而盐浓度高于0.4 mol/L则呈减少的趋势,但粒径均大于1.1μm;而当ELPs末端融合木聚糖酶和1,3-丙二醇氧化还原酶后,其自组装形成的微球粒径急剧减小,约为游离ELPs的1/10,分别为151.0 nm和174.2 nm。导致这种现象的原因可能是酶分子和ELPs通过静电引力相互作用后,酶分子的空间位阻妨碍了ELPs分子的聚集。     

茉莉酸和双氧水对斜生褐孔菌一株黑色素缺失突变体产生酚类化合物的影响

黑色素合成是桦褐孔菌Phaeoporus obliquus次生代谢过程中的一个重要事件。黑色素的过多合成势必影响其他酚类的积累。本文以茉莉酸和双氧水为调控因子,开展了该突变体深层发酵过程酚类化合物积累研究。正常培养条件下,每升发酵液积累酚类化合物最高可达317.45mg/L,其中包括苯甲酸衍生物(45.3mg/L),黄酮苷元(74.4mg/L),黄酮苷(160mg/L)以及硬毛素类化合物(18.5mg/L)。加入茉莉酸后,总酚最大值增加到620.45mg/L,其中苯甲酸衍生物增加到230.26mg/L,黄酮苷元增加到166.86mg/L,而黄酮苷和硬毛素类化合物却分别降低到98.5mg/L和7.1mg,双氧水的加入没有提高总酚的最大积累(315.69mg/L)。因此,与野生型相比,黑色素缺失使总酚的积累有所增加,随茉莉L酸的加入总酚积累最大值增加近一倍。虽然双氧水的加入提高了苯丙氨酸裂解酶活性,但总酚的积累却没有增加,这可能是由于一部分酚类化合物在抗双氧水氧化胁迫过程中被消耗。     

Preparations of immobilized lysozyme with reversibly soluble polymer for hydrolysis of microbial cells

Hen egg white lysozyme was immobilized by carbodiimide method to form amide bonds with a polymer (AS-L) showing reversibly soluble-insoluble characteristics with pH change. The immobilized enzyme (LY-AS) was soluble above pH 6 and precipitate below pH 4.5, offering advantages in that it can carry out hydrolysis of microbial cells in a soluble form yet be recovered after precipitation at low pH. The maximum specific activity of LY-AS was 66% of that of free lysozyme with M. lysodeikticus cells as substrate, which is much higher than the values reported in the literature using water-insoluble materials as carriers. The effects of pH and temperature on the activity of LY-AS were studied and compared with those of free lysozyme. With repeated pH cycles between 6.6 and 4.5, the operation half-life of immobilized enzyme activity was nine cycles. Repeated batch lysis of microbial cells could be carried out with intermittent enzyme precipitation and recovery steps. In such an operation the insoluble residual cells should be recovered together with the immobilized enzyme to minimize enzyme loss arising from adsorption to cells.     

Mechanism of selective stabilization of extrinsic polypeptides in PSII particles by glycinebetaine

Ｔｈｒｅｅｅｘｔｒｉｎｓｉｃｐｏｌｙｐｅｐｔｉｄｅｓ,ｗｈｉｃｈａｒｅｌｏｃａｔｅｄｏｎｔｈｅｉｎｎｅｒｓｕｒｆａｃｅｏｆｔｈｙｌａｋｏｉｄｍｅｍｂｒａｎｅ,ｐｌａｙｅｓｓｅｎｔｉａｌｒｏｌｅｓｉｎｍａｉｎｔａｉｎｉｎｇｔｈｅｐｈｏｔｏｓｙｎｔｈｅｔｉｃｅｖｏｌｕｔｉｏｎｏｆｏｘｙｇｅｎ.Ｔｈｅｙａｒｅｇｅｎｅｒａｌｌｙｒｅｃｏｇｎｉｚｅｄａｓｌｉａｂｌｅｃｏｍｐｏｎｅｎｔｓｏｆｔｈｅｐｈｏｔｏｓｙｎｔｈｅｔｉｃａｐｐａｒａｔｕｓ,ａｎｄｃａｎｂｅｒｅｌｅａｓｅｄｂｙａｖａｒｉｅｔｙｏｆｐｈｙｓｉｃ…     

Separation,antitumor activities,and encapsulation of polypeptide from Chlorella pyrenoidosa

Chlorella pyrenoidosa is a unicellular green algae and has been a popular foodstuff worldwide. However, no reports on the antitumor peptides from such a microalgae are available in the literature. In this study, using low‐temperature high‐pressure extraction, enzymatic hydrolysis, ion exchange, and gel filtration chromatography, we separated a polypeptide that exhibited inhibitory activity on human liver cancer HepG2 cells, and named the polypeptide CPAP (C. pyrenoidosa antitumor polypeptide). Furthermore, the micro‐ and nanoencapsulation of CPAP were investigated by using two methods: complex coacervation and ionotropic gelation. The in vitro release tests revealed that CPAP was well preserved against gastric enzymatic degradation after micro/nanoencapsulation and the slowly controlled release in the intestine could be potentially achieved. These results suggest that CPAP may be a useful ingredient in food, nutraceutical, and pharmaceutical applications. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:681–687, 2013     

植物的低温蛋白

综述了与植物耐冻性有关的一些植物内源蛋白质或多肽 ,包括低温防护蛋白、抗冻蛋白、植物脱水素、膜关联耐冻性多肽蛋白质。结果表明 ,植物的耐冻性与其低温蛋白 (cold induced proteins)有着密切的关系 ,并指出了抗冻蛋白行使功能的两种可能的作用方式。同时 ,耐冻性与除低温外的其它环境胁迫因子的植物抗性如抗干旱、抗病虫、高盐耐性、乙烯耐性等密切相关     

Biochemical correlates to cortical dysplasia,gliosis, and astrocytoma infiltration in human epileptogenic cortex

The study provides detailed biochemical correlates to the common histopathological diagnoses in epilepsy. A dot immunobinding procedure was used for quantification of NSE, GFA, S-100, NCAM, NF 68 and NF 200. The material consisted of samples from 48 patients either selected for surgical treatment of partial epilepsy or for disorders not related to epilepsy. The histopthological diagnosis of the epileptic cases was: MCD (mild cortical dysplasia, microdysgenesis), gliosis, astrocytoma, ganglioglioma, oligodendroglioma and single cases. The concentration in non-epileptic white matter, in per cent of that in grey matter was: NSE, 85; GFA, 175; S-100, 117; NCAM, 43; NF 68,227 and NF 200, 173. The concentration of NSE as well as of GFA was close to normal in the specimens of the MCD and gliosis groups and of one subgroup of the astrocytomas. There was a striking inverse relationship of the GFA vs the NSE concentrations in the whole material. The concentrations of S-100 showed no such inverse relationship to NSE levels. In all the epileptic groups, total NCAM was lower than 50% of that of the non-epileptic group. The mean NF 68 and NF 200 concentration in the gliosis and astrocytoma groups was 75% of the of the non-epileptic group while the corresponding value for the MCD group was 50%. There was a positive correlation of immunochemically determined GFA and the histopathological gliosis score in the samples of epileptogenic cortex. There was no correlation between the concentration of GFA in the samples and the duration of epilepsy. The concentration of neuronal markers was relatively unaffected in the cortex of most patients with epilepsy related to MCD, gliosis and even to astrocytoma infiltration, even after years of seizures.Special issue dedicated to Dr. Claude Baxter.     

Biochemical composition and photosynthetic activity of chloroplasts from Striga hermonthica and Striga aspera, root parasites of field-grown cereals

Striga hermonthica (Del.) Benth. and Striga aspera (Willd.) Benth. are root parasites causing dramatic losses in field-grown cereals in semi-and tropics. Being achlorophyllous and obligate parasites during their underground development, upon emergence from the soil, they become green leafy plants; but, despite the presence of chlorophyll, they exhibit only low rates of photosynthesis. To investigate if deficiency in the photosynthetic apparatus could account for the low rates of photosynthesis, chloroplasts were isolated from S. hermonthica parasitizing sorghum [Sorghum bicolor (L.) Moench cv. Tiemarifing] and from S. aspera parasitizing maize ( Zea mays L. cv. Tiémantié ) grown under greenhouse conditions or in their natural surroundings. Isolated chloroplasts exhibited the characteristics of chloroplasts from C₃ plants but displayed low levels of chlorophyll and polar lipids, while the protein content was less reduced. Main changes occurred in polar lipid composition, with decreases in monogalactosyldiacylglycerol and digalactosyldiacylglycerol. All polar lipids showed a decrease in the degree of unsaturation of fatly acids. All these changes were particularly pronounced in chloroplasts from plants that experienced heavy drought in Africa. On a chlorophyll basis, chloroplasts did not display a dramatic decrease in photosynthetic activities. These results are discussed in relation to parasitism and drought adaptation.     

A preparative method for the isolation of the 43 kDa, 47 kDa and the D1-D2-Cyt b 559 species directly from thylakoid membranes

Using the non-ionic detergent dodecyl--D-maltoside we have developed a preparative method for the isolation of the 43 kDa, 47 kDa and D₁-D₂-Cyt b ₅₅₉ species directly from thylakoid membranes. In contrast to previous procedures the photosynthetic membrane was exposed only to one mild detergent and that resulted in more stable preparations. The isolated species were examined spectroscopically and it was found that even under these mild conditions the D₁-D₂-Cyt b ₅₅₉ did not retain the primary quinone Q_A.Abbreviations PS II Photosystem II - CF Coupling Factor - LHC Light Harvesting Complex - MES 4-morpholine-ethanesulfonic acid - BIS-TRIS 2-[Bis (2-hydroxyethyl) amino]-2-hydroxymethyl-propane-1,3-diol - SMN 0.4 M sucrose/50 mM MES (pH6)/10 mM NaCl