抗生素和干扰素

860582 acidocin-A的分离和鉴定以及细菌素基因的克隆[英]/Kanatani,K…∥Appl.Environ.Microbiol.-1995,61(3).-1061～1067[译自DBA,1995,14(11),95-06273] 通过硫酸铵沉淀、离子交换色谱和反相色谱法将产生于嗜酸乳杆菌TK9201的细菌素acidocin-A纯化至均质。该物质的分子量为6500。测定了其N-     

细菌素Paracin 1.7的纯化与性质

[目的]分离纯化(Lactobacillus paracasei)HD1.7所产生的细菌素并分析其特性.[方法]细菌素Paracin1.7的纯化采用色谱技术,其分子量检测采用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE),利用琼脂扩散法测定细菌素活力.[结果]Paracin 1.7分离于我国传统发酵食品酸菜发酵液中,其产生菌为副干酪乳杆菌.Paracin 1.7可以抑制其它微生物的生长,为细菌素.该菌在稳定期可产生大量Paracin 1.7.经过阳离子交换层析、凝胶过滤层析以及高效液相色谱(HPLC),对该细菌素进行了初步纯化,并经Tricine-SDS-PAGE检测其分子量大约为11 kDa.Paracin 1.7抑菌谱较广,其抑菌范围包括Proteus,Bacillus,Enterobacter,Staphylococcus,Escherichia,Lactobacillus,Microccus,Pseudomonas,Salmonella,Saccharomyces,其中有些为食品源致病菌.该细菌素在酸性及高温下稳定,对几种蛋白质酶敏感.该细菌素对敏感菌株的作用方式为抑菌.在4℃保存4个月后,Paraein 1.7的抑菌活性保持稳定.[结论]基于细菌素Paracin 1.7的性质,该细菌素可用作食品防腐剂.     

人血清白蛋白纯化技术研究进展

本文综述了国内外关于血浆人血清白蛋白（pHSA）和基因重组人血清白蛋白（rHSA）分离纯化的进展和发展趋势。以冷乙醇沉淀为主的pHSA分离方法仍是目前多数工业采用的工艺,但近年来发展的离子交换色谱、凝胶过滤色谱、亲和色谱等多种色谱分离技术具有自动化程度高、生产周期短、更符合GMP等特点,正在逐步取代传统的冷乙醇沉淀法。当前用基因工程技术表达的人血清白蛋白对分离纯化提出了新的挑战。为获得高纯度、安全、稳定的产品,各种色谱分离技术得到更多的应用,其中扩张床离子交换色谱可以省去离心、过滤等传统固液分离操作。尽管rHSA的纯化技术已有一定的发展,但纯化过程仍需进一步优化以提高产品纯度及收率。     

细菌素Paracin 1.7的纯化与性质研究

摘要：【目的】分离纯化（Lactobacillus paracasei）HD1.7所产生的细菌素并分析其特性。【方法】细菌素Paracin 1.7的纯化采用色谱技术,其分子量检测采用十二烷基磺酸钠－聚丙烯酰胺凝胶电泳（SDS-PAGE）,利用琼脂扩散法测定细菌素活力。【结果】Paracin 1.7分离于我国传统发酵食品酸菜发酵液中,其产生菌为副干酪乳杆菌。 Paracin 1.7可以抑制其它微生物的生长,为细菌素。该菌在稳定期可产生大量Paracin 1.7。经过阳离子交换层析、凝胶过滤层析以及高效液相色谱（HPLC）,对该细菌素进行了初步纯化,并经Tricine-SDS-PAGE检测其分子量大约为11 kDa。Paracin 1.7抑菌谱较广,其抑菌范围包括Proteus, Bacillus, Enterobacter, Staphylococcus, Escherichia, Lactobacillus, Microccus, Pseudomonas, Salmonella, Saccharomyces,其中有些为食品源致病菌。该细菌素在酸性及高温下稳定,对几种蛋白质酶敏感。该细菌素对敏感菌株的作用方式为抑菌。在4oC保存4个月后,Paracin 1.7的抑菌活性保持稳定。【结论】基于细菌素Paracin 1.7的性质,该细菌素可用作食品防腐剂。     

细菌素异源表达研究进展

细菌素有望成为抗生素替代品和新型绿色生物兽药,但细菌素野生菌种产量低,细菌素分离纯化困难,为了提高细菌素产量,许多研究者进行了细菌素异源表达研究,本文就细菌素异源表达进行综述。     

一株布氏乳杆菌所产类细菌素的初步纯化与部分特性

从分离自内蒙古传统乳制品的67株乳酸菌中筛选得到一株产生类细菌素的布氏乳杆菌KLDS1.0364, 对其所产类细菌素进行初步分离纯化, 同时研究其所产类细菌素的生物学特性。KLDS1.0364无细胞发酵上清液经阳离子交换树脂纯化后, 采用tricine-SDS-PAGE测定类细菌素分子量, 并测定了类细菌素的部分特性。KLDS1.0364产生的类细菌素分子量约为21.6kD, 对热和pH值稳定, 可被多种蛋白酶失活, 不能被过氧化氢酶和α-淀粉酶失活。KLDS1.0364产生的类细菌素的作用方式是杀菌, 且抑菌谱广, 可抑制多种革兰氏阳性菌、革兰氏阴性菌和真菌。     

离子交换法与氨基酸的分离纯化

离子交换法广泛用于氨基酸的分离纯化。文章综述了离子交换法与氨基酸的分离纯化,包括分离纯化单一氨基酸及对氨基酸混合物进行分组分离纯化。     

血链球菌细菌素抑菌活性研究

检测分离纯化的血链球菌细菌素对牙周可疑致病菌的抑制作用。通过羟基磷灰石(HA)柱层析、SephadexG-150凝胶柱层析、中空纤维柱超滤脱盐、浓缩纯化提取血链球菌细菌素,以具核梭杆菌为指示菌,洞平板法检测细菌素的抑菌活性。经HA柱层析得到4个相互分离的组分,经洞平板法检测,第Ⅱ峰的蛋白具有抑菌活性,冻干后得纯化后的细菌素,终产率为0.082%;1mg/mL的细菌素溶液可形成17mm的抑菌环,最小抑菌浓度为62.5μg/mL。血链球菌细菌素对牙周可疑致病菌具有较强的拮抗作用。     

一株布氏乳杆菌所产类细菌素的初步纯化与部分特性

从分离自内蒙古传统乳制品的67株乳酸菌中筛选得到一株产生类细菌素的布氏乳杆菌KLDS1.0364,对其所产类细菌素进行初步分离纯化,同时研究其所产类细菌素的生物学特性.KLDS1.0364无细胞发酵上清液经阳离子交换树脂纯化后,采用tricine-SDS-PAGE测定类细菌素分子量,并测定了类细菌素的部分特性.KLDS1.0364产生的类细菌素分子量约为21.6kD,对热和pH值稳定,可被多种蛋白酶失活,不能被过氧化氢酶和α-淀粉酶失活.KLDS1.0364产生的类细菌素的作用方式是杀菌,且抑菌谱广,可抑制多种革兰氏阳性菌、革兰氏阴性菌和真菌.     

纯化小麦α-淀粉酶的亲和层析法

纯化α-淀粉酶有多种方法。Mac Gregor等以及Kruger和Tkachuk分别应用羧甲基纤锥素离子交换层析与丙酮分部、糖元沉淀和离子交换层析,各自从大麦及春小麦分离了α-淀粉酶。Tkachuk又发展了以α-环化糊精为配基的α-淀粉酶分离纯化的亲和层析法。由于亲和层析法简便、快速、高效,因而获得了广泛的应用。我们以β-环化糊精(β-Cyclodextrin)为配基,分离纯化了小麦α-淀粉酶。现介绍如下。     

Method for rapid purification of class IIa bacteriocins and comparison of their activities

A three-step method was developed for the purification of mesentericin Y105 (60% yield) from the culture supernatant of Leuconostoc mesenteroides Y105. The same procedure was successfully applied to the purification of five other anti-Listeria bacteriocins identified by mass spectrometry. Specific activities of the purified bacteriocins were compared.     

Commercial ampholytes used for isoelectric focusing may interfere with bioactivity based purification of antimicrobial peptides

BioRad's Rotofor system has been frequently used for the purification of proteins and smaller peptides such as bacteriocins. In this study, we report that some commercially available ampholytes used with the Rotofor isoelectric focusing system possess antimicrobial activity, which may interfere with the purification of bacteriocins and bacteriocin-like substances.     

Method for Rapid Purification of Class IIa Bacteriocins and Comparison of Their Activities

A three-step method was developed for the purification of mesentericin Y105 (60% yield) from the culture supernatant of Leuconostoc mesenteroides Y105. The same procedure was successfully applied to the purification of five other anti-Listeria bacteriocins identified by mass spectrometry. Specific activities of the purified bacteriocins were compared.     

Bacteriocin Production and Different Strategies for Their Recovery and Purification

Bacteriocins from lactic acid bacteria (LAB) are a diverse group of antimicrobial proteins/peptides, offering potential as biopreservatives, and exhibit a broad spectrum of antimicrobial activity at low concentrations along with thermal as well as pH stability in foods. High bacteriocin production usually occurs in complex media. However, such media are expensive for an economical production process. For effective use of bacteriocins as food biopreservatives, there is a need to have heat-stable wide spectrum bacteriocins produced with high-specific activity in food-grade medium. The main hurdles concerning the application of bacteriocins as food biopreservatives is their low yield in food-grade medium and time-consuming, expensive purification processes, which are suitable at laboratory scale but not at industrial scale. So, the present review focuses on the bacteriocins production using complex and food-grade media, which mainly emphasizes on the bacteriocin producer strains, media used, different production systems used and effect of different fermentation conditions on the bacteriocin production. In addition, this review emphasizes the purification processes designed for efficient recovery of bacteriocins at small and large scale.     

Bacteriocins: criteria,classification, characteristics,methods of detection

The review on bacteriocins of Gram negative and Gram positive bacteria. Criteria making it possible to regard antagonistic substances as bateriocins or bacteriocin-like substances and on their classification are presented. Examples of bacteriocins naming depending on the taxonomic position of the producer culture are given. Information on the physico-chemical and biological properties of bacteriocins and their purification is presented as well as on detection tools of bacteriocins in microorganisms and evaluation of the producer activity of the bacteriological culture.     

Rapid Two-Step Procedure for Large-Scale Purification of Pediocin-Like Bacteriocins and Other Cationic Antimicrobial Peptides from Complex Culture Medium

A rapid and simple two-step procedure suitable for both small- and large-scale purification of pediocin-like bacteriocins and other cationic peptides has been developed. In the first step, the bacterial culture was applied directly on a cation-exchange column (1-ml cation exchanger per 100-ml cell culture). Bacteria and anionic compounds passed through the column, and cationic bacteriocins were subsequently eluted with 1 M NaCl. In the second step, the bacteriocin fraction was applied on a low-pressure, reverse-phase column and the bacteriocins were detected as major optical density peaks upon elution with propanol. More than 80% of the activity that was initially in the culture supernatant was recovered in both purification steps, and the final bacteriocin preparation was more than 90% pure as judged by analytical reverse-phase chromatography and capillary electrophoresis.     

Rapid two-step procedure for large-scale purification of pediocin-like bacteriocins and other cationic antimicrobial peptides from complex culture medium

A rapid and simple two-step procedure suitable for both small- and large-scale purification of pediocin-like bacteriocins and other cationic peptides has been developed. In the first step, the bacterial culture was applied directly on a cation-exchange column (1-ml cation exchanger per 100-ml cell culture). Bacteria and anionic compounds passed through the column, and cationic bacteriocins were subsequently eluted with 1 M NaCl. In the second step, the bacteriocin fraction was applied on a low-pressure, reverse-phase column and the bacteriocins were detected as major optical density peaks upon elution with propanol. More than 80% of the activity that was initially in the culture supernatant was recovered in both purification steps, and the final bacteriocin preparation was more than 90% pure as judged by analytical reverse-phase chromatography and capillary electrophoresis.     

重组人粒细胞-巨噬细胞集落刺激因子复性和纯化方法的比较

比较重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)在用疏水色谱(HIC)、离子交换色谱(IEC)和体积排阻色谱(SEC)3种液相色谱进行复性和纯化,选择较好的复性和纯化方法.通过对比rhGM-CSF在液相色谱上复性和纯化的主要指标,包括比活、纯度和质量回收率,结果发现采用HIC和IEC对rhGM-CSF进行复性和纯化时,其比活可以达到国家标准,纯度和质量回收率也比较高,而采用SEC,其比活、纯度和质量回收率远低于HIC和IEC.     

HPLC purification and re‐evaluation of chemical identity of two circular bacteriocins,gassericin A and reutericin 6

Aim: The study aimed for the complete purification and recharacterization of the highly hydrophobic circular bacteriocins, gassericin A and reutericin 6. Methods and Results: Gassericin A and reutericin 6 were purified to homogeneity using previously described method and reverse‐phase HPLC with an octyl column and eluents of aqueous acetonitrile and 2‐propanol. Mass analysis, N‐terminal sequencing and bacteriocin assay of the HPLC‐purified bacteriocins showed the two bacteriocins had identical seamless circular structures with the same m/z value (5651) of [M + H]⁺ and both had the same specific activity. d/l‐ amino acid composition analysis using two distinct methods with the chiral fluorescent derivatization reagents (+)‐1‐(9‐fluorenyl)ethyl chloroformate and o‐phthalaldehyde/N‐acetyl‐l ‐cystein revealed neither gassericin A nor reutericin 6 contained d ‐alanine residues contrary to our previous results. Conclusion: Purified gassericin A and reutericin 6 are chemically identical circular molecules containing no d ‐alanine residues. Significance and Impact of the Study: The HPLC conditions developed in this study will facilitate advanced purification and correct characterization of other highly hydrophobic bacteriocins.     

Concentration and purification of rubella virus using monolithic chromatographic support

The production of economically acceptable viral vaccines of high quality requires simple and efficient methods for purification and concentration of viral particles. Ion-exchange chromatography (IEC) has become one of commonly used methods for large-scale downstream purification of viruses. Viruses possess different biological and/or biochemical properties and therefore IEC conditions must be established specifically for each virus. Live attenuated rubella virus vaccines have been manufactured and successfully used widely to protect people from rubella and congenital rubella syndrome for almost 40 years. The aim of this study was to search for an efficient method for concentration and purification of rubella virus using IEC. The selected operating conditions using quaternary amine monolithic supports enabled highly efficient binding, purification and concentration of rubella virus from complex biological suspension without additional procedures. Eluted viral particles maintained their infectivity and viral recovery was almost 100%. At the same time, viral preparation was successfully depleted from host cell protein and DNA. This work indicates the possibility of using monoliths to improve the rubella virus yields in productions where high virus titers during cultivation can hardly be achieved.