克拉维酸发酵液中甘油含量的测定

为寻求一种简便易行的克拉维酸发酵液中甘油含量的测定方法,以化学测定方法一高碘酸氧化法对克拉维酸发酵液进行测定。从精密度和回收率实验结果看,该方法可以较精确地测定出发酵液中甘油的含量,平均相对偏差小于5％,适合用来检测克拉维酸发酵过程中甘油的代谢情况。     

影响克拉维酸生物合成的氨基酸

发酵液的氨基酸分析显示,谷氨酸,精氨酸,天门冬氨酸,丙氨酸易被棒状链霉菌利用,发酵培养基中添加上述氨基酸后,谷氨酸,精氨酸有利于克拉维酸的生物合成,适时添加谷氨酸,精氨酸可分别提高克拉维酸的产量约25%和12%;而蛋氨酸,半胱氨酸含S氨基酸对克拉维酸生物合成不利,不同来源的黄豆粉作发酵培养基氮源,因其组成中某些氨基酸含量的差异。可使克拉维酸的产量相差百分之十几。     

克拉维酸生产中废乙酸乙酯的紫外扫描评价方法及树脂处理

建立克拉维酸生产中废乙酸乙酯的快捷评价方法。采用紫外吸收扫描,以吸收面积作为评价指标,全面评价废乙酸乙酯中的杂质残留。选用不同的树脂吸附处理废乙酸乙酯。结果表明:废乙酸乙酯经FPA90Cl树脂吸附处理后,紫外吸收面积最小,仅有225.601±5.499,残留的杂质最少。经条件优化后,批处理量为60 m3废乙酸乙酯的树脂用于克拉维酸生产,产品质量与新乙酸乙酯生产的产品质量相近。使用树脂处理废乙酸乙酯可减轻乙酸乙酯蒸馏回收和环保的压力,降低生产成本,具有良好的经济效益和环境效益。     

克拉维酸产生菌的优化培养

为提高克拉维酸产量,通过摇瓶实验考查了多种氮,碳源及无机盐对克拉维酸产量的影响,并对产生菌的部分性能进行了考查,试验表明实验中所用棒状链霉菌（Strepto-myces elavuligerus）合成克拉维酸的最佳氮,碳源及无机盐是黄豆粉,甘油,KH2PO4,MgSO4,其最佳培养基配比（%）分别为2.8、1.8、0.04、0.05。为保证克拉维酸高产,稳产,产生菌斜面冷藏时间不宜超过1个月,斜面传代次数不宜超过4代,沙土,20%甘油溶液,冷冻干燥管均可用于产生菌的孢子保藏。     

阿莫西林克拉维酸钾或头孢他啶联合布地奈德对肺炎患儿的疗效

目的 探讨阿莫西林克拉维酸钾或头孢他啶联合布地奈德对肺炎患儿的治疗效果及对肠道微生物的影响。 方法 选择2016年9月至2018年12月我院儿科收治的90例肺炎患儿为研究对象,入选患儿随机分为对照组(n=45)和研究组(n=45)。另外选取本院同期健康体检的40例儿童为健康组。对照组患儿接受头孢他啶联合布地奈德疗法,研究组患儿采用阿莫西林克拉维酸钾联合布地奈德疗法。连续治疗7 d后,比较两组患儿临床疗效、临床症状(发热、咳嗽、啰音及扁桃体充血)消失时间及不良反应,同时比较各组患儿肠道微生物(乳杆菌、双歧杆菌、真杆菌、肠杆菌及肠球菌)变化情况。 结果 研究组患儿总有效率为95.56%,对照组总有效率为91.11%,两组比较差异无统计学意义(χ2=0.714,P=0.398)。研究组患儿发热、咳嗽、啰音及扁桃体充血消失时间与对照组比较差异无统计学意义(均P>0.05)。治疗前,研究组和对照组患儿肠道菌群数量与健康组比较差异无统计学意义(均P>0.05)。治疗后,研究组和对照组患儿肠道菌群数量与健康组比较差异有统计学意义(均P0.05)。研究组(8.89%)和对照组(13.33%)总不良反应率比较差异无统计学意义(P>0.05)。 结论 阿莫西林克拉维酸钾联合布地奈德治疗小儿肺炎疗效显著,能有效缩短患儿临床症状持续时间,其疗效与头孢他啶联合布地奈德疗效相当,且药物副作用较小,但对肠道菌群平衡有破坏作用。因此,扶植肠道菌群可以作为综合治疗的一部分。     

正交试验法探讨单酸降解纤维素的最佳条件

正交试验法探讨了乙酸、草酸、盐酸、硫酸等单酸降解植物纤维素的最佳条件。研究结果表明,乙酸降解纤维素时,影响因素的主次顺序为：A＞B＞C,最佳组合为A3B1C2。即乙酸浓度为30％,温度40℃,反应1．5h为最佳条件,验证性试验结果表明,单用乙醇能使纤维素降解成葡萄糖的转化率达42．2％。而草酸影响因素的主次则为A＞C＞B,最佳组合为A2C1B2。即草酸度浓度0％,70℃酸解1h为最佳条件。在此条件下,转化率可达40．8％。对于盐酸影响因素的主次为A＞B＞C,最佳组合为A3B3C1。即在20％盐酸,100℃酸解1h的条件下,转化率可达42．8％。而硫酸影响因素的主次顺序为B＞A＞C,最佳组合为A3B3C1。即在40％硫酸,100℃反应1h的最佳条件下能使转化率达40．9％。研究还发现,乙酸、草酸、盐酸、硫酸降解纤维素特性不同。乙酸、草酸同属有机酸,但因二者的酸性、挥发性、溶解性等不同,致使温度、时间两因素对纤维降解的影响相关很大。盐酸、硫酸无机强酸,都要求高温短时,但随着浓度的升高,转化度呈下降趋势,且二者的下降幅度不同,硫酸大于盐酸。     

阿莫西林克拉维酸钾联合阿奇霉素治疗社区获得性肺炎的临床疗效研究

目的：探讨阿莫西林克拉维酸钾联合阿奇霉素治疗社区获得性肺炎（CAP）的临床疗效。方法：选取于2010年1月-2012年1月收治的CAP患者145例,采用随机数字表法将其分为两组,其中治疗组73例,对照组72例。治疗纽给予阿莫西林克拉维酸钾联合阿奇霉素治疗,对照组只单纯给予应用阿莫西林克拉维酸钾。两组疗程均为7～14d。于治疗后比较两组的临床疗效。结果：（1）治疗组治愈、显效、有效、无效分别为43例（58．9％）、16例（21．9％）、8例（11．0％）、6例（8．2％）;对照组分别为20例（27．8％）、26例（36．1％）、14例（19．4％）、12例（16．7％）。两组疗效比较,差异有统计学意义（P〈0．05）。（2）治疗组胃肠道反应、静脉滴注局部疼痛、皮疹的发生分别为6例（8．2％）、7例（9．6％）、3例（4．1％）,总不良反应发生率为16例（21．9％）;对照组分别为5例（6．9％）、6例（8.3％）、3例（4．2％）、14例（19．4％）。两组不良反应发生率比较,差异无统计学意义（P〉0．05）。结论：阿莫西林克拉维酸钾联合阿奇霉素治疗CAP疗效好。     

阿莫西林克拉维酸钾联合阿奇霉素治疗社区获得性肺炎的临床疗效研究

目的:探讨阿莫西林克拉维酸钾联合阿奇霉素治疗社区获得性肺炎(CAP)的临床疗效。方法:选取于2010年1月-2012年1月收治的CAP患者145例,采用随机数字表法将其分为两组,其中治疗组73例,对照组72例。治疗组给予阿莫西林克拉维酸钾联合阿奇霉素治疗,对照组只单纯给予应用阿莫西林克拉维酸钾。两组疗程均为7~14 d。于治疗后比较两组的临床疗效。结果:(1)治疗组治愈、显效、有效、无效分别为43例(58.9%)、16例(21.9%)、8例(11.0%)、6例(8.2%);对照组分别为20例(27.8%)、26例(36.1%)、14例(19.4%)、12例(16.7%)。两组疗效比较,差异有统计学意义(P0.05)。(2)治疗组胃肠道反应、静脉滴注局部疼痛、皮疹的发生分别为6例(8.2%)、7例(9.6%)、3例(4.1%),总不良反应发生率为16例(21.9%);对照组分别为5例(6.9%)、6例(8.3%)、3例(4.2%)、14例(19.4%)。两组不良反应发生率比较,差异无统计学意义(P0.05)。结论:阿莫西林克拉维酸钾联合阿奇霉素治疗CAP疗效好。     

混酸法降解植物纤维素技术研究

以单酸正交试验结果为基础,对二酸混合法和三酸混合法降解植物纤维素的最佳工艺条件进行了探讨。二酸混合法的结果表明,酸的添加顺序严重影响纤维素向葡萄糖的转化。强酸与弱酸同时混用均使葡萄糖的转化率降低。但盐酸与乙酸的混用效果则因所选最佳条件的不同而不同,虽然硫酸与盐酸均为无机强酸,与乙酸混用时添加的顺序亦一样即先弱酸后强酸,但其结果差别甚大,前者使葡萄糖的转化率降低,后者则使葡萄糖的转化率由40.9%锰增到62.7%。三酸混合法的结果表明,在本研究所选用的四种酸中,无论是哪三种酸同时混用,均使葡萄糖的转化率降低;当按一定顺序添加时,其结果亦不如二酸混合法,结论：在用多种酸降解植物纤维素时,一定要按一定的顺序添加,最好用两种酸即可。     

聚对苯二甲酸乙二醇酯降解酶的研究进展

聚对苯二甲酸乙二醇酯(Polyethylene terephthalate,PET)因其良好的耐用性和可塑性,已在世界范围内的工业领域和日常生活中得到广泛应用。目前自然环境中大量PET使用废弃物的积累和迁移给全球生态系统带来了严重负担,因此PET的降解问题已成为全球性的热点问题。微生物酶降解法目前被认为是一种理想绿色PET降解方法,有希望应用于大规模降解PET废弃物降解处理。传统的PET降解酶主要包括脂肪酶、酯酶和角质酶等,但这些酶的PET降解活性相对不高。近期科学家从Ideonella sakaiensis细菌中分离了一种新型水解酶PETase,能够特异性高效降解PET。本文从结构生物学角度对多种PET降解酶进行梳理,重点总结了新近发现的PETase催化机制,为发展改造更有效的PET降解酶提供理论依据。     

Clavulanic acid: a review

Natural antibiotics are almost universal secondary metabolites, not essential for the growth of the producing organisms generally produced at low growth rates or after growth has ceased. Clavulanic acid (CA), a naturally occurring powerful inhibitor of bacterial beta-lactamases is a major beta-lactam antibiotic produced by organism Streptomyces clavuligerus and is active against a wide spectrum of Gram-positive and Gram-negative bacteria. The review discusses the biosynthetic pathway, fermentative production, downstream processing and applications of CA.     

Solution Stability and Degradation Pathway of Deoxyribonucleoside Phosphoramidites in Acetonitrile

The impuritiy profiles of acetonitrile solutions of the four standard O‐cyanoethyl‐N,N‐diisopropyl‐phosphoramidites of 5′‐O‐dimethoxytrityl (DMT) protected deoxyribonucleosides (dG^ib, dA^bz, dC^bz, T) were analyzed by HPLC‐MS. The solution stability of the phosphoramidites decreases in the order T, dC>dA>dG. After five weeks storage under inert gas atmosphere the amidite purity was reduced by 2% (T, dC), 6% (dA), and 39% (dG), respectively. The main degradation pathways involve hydrolysis, elimination of acrylonitrile and autocatalytic acrylonitrile‐induced formation of cyanoethyl phosphonoamidates. Consequently, the rate of degradation is reduced by reducing the water concentration in solution with molecular sieves and by lowering the amidite concentration. Acid‐catalyzed hydrolysis could also be reduced by addition of small amounts of base.     

Biosynthesis and Molecular Genetics of Clavulanic Acid

The biosynthesis of clavulanic acid and related clavam metabolites is only now being elucidated. Understanding of this pathway has resulted from a combination of both biochemical studies of purified biosynthetic enzymes, and molecular genetic studies of the genes encoding these enzymes. Clavulanic acid biosynthesis has been most thoroughly investigated in Streptomyces clavuligerus where the biosynthetic gene cluster resides immediately adjacent to the cluster of cephamycin biosynthetic genes. A minimum of eight structural genes have been implicated in clavulanic acid biosynthesis, although more are probably involved. While details of the early and late steps of the pathway remain unclear, synthesis proceeds from arginine and pyruvate, as the most likely primary metabolic precursors, through the monocyclic -lactam intermediate, proclavaminic acid, to the bicyclic intermediate, clavaminic acid, which is a branch point leading either to clavulanic acid or the other clavams. Conversion of clavaminic acid to clavulanic acid requires side chain modfication as well as inversion of ring stereochemistry. This stereochemical change occurs coincident with acquisition of the -lactamase inhibitory activity which gives clavulanic acid its therapeutic and commercial importance. In contrast, the other clavam metabolites all arise from clavaminic acid with retention of configuration and lack -lactamase inhibitory activity.     

Hyaluronic Acid Degradation by Ascorbic Acid and Influence of Iron

The effects of ascorbic acid, iron and ADP on hyaluronic acid, a compound present in inflamed joints, were investigated in an in vitro system. Ascorbic acid induces degradation of hyaluronic acid which increased in the presence of FeCl, and which is additionally stimulated by ADP chelated ferric ions. The hyaluronic acid degrading reactions induced by the Fe-III/ADP/ascorbic acid system were inhibited by catalase and formate to various extents whereas the presence of superoxide dismutase did not exert any inhibitory effect. Desferrioxamine, a specific iron chelator, completely inhibited hyaluronic acid depolymerisation by ascorbic acid as well as in combination with FeCl₃ or FeCl₃/ADP, respectively. We suggest that the ultimate hyaluronic acid degrading species is OH', generated via the Fe-III/ADP catalysed Haber Weiss reaction. There is also an indication for the involvement of perferryl or/and ferryl species in the degradation process.     

Hyaluronic Acid Degradation by Ascorbic Acid and Influence of Iron

The effects of ascorbic acid, iron and ADP on hyaluronic acid, a compound present in inflamed joints, were investigated in an in vitro system. Ascorbic acid induces degradation of hyaluronic acid which increased in the presence of FeCl, and which is additionally stimulated by ADP chelated ferric ions. The hyaluronic acid degrading reactions induced by the Fe-III/ADP/ascorbic acid system were inhibited by catalase and formate to various extents whereas the presence of superoxide dismutase did not exert any inhibitory effect. Desferrioxamine, a specific iron chelator, completely inhibited hyaluronic acid depolymerisation by ascorbic acid as well as in combination with FeCl₃ or FeCl₃/ADP, respectively. We suggest that the ultimate hyaluronic acid degrading species is OH', generated via the Fe-III/ADP catalysed Haber Weiss reaction. There is also an indication for the involvement of perferryl or/and ferryl species in the degradation process.     

纳米双相磷酸钙陶瓷支架材料肌内降解性能的实验研究

目的:纳米双相磷酸钙陶瓷(Biphasic calcium phosphate nanocomposite,NanoBCP)支架是一种新型支架材料,具有三维立体多孔结构,孔隙率可达60%～80%。本研究观察了纳米双相磷酸钙陶瓷肌内降解情况。方法:将NanoBCP制备为5mm×5mm×1.5mm大小各8块的支架植入SD大鼠腿部肌袋内,相同孔径、孔隙率的羟基磷灰石(Hydroxyapatite,HA)及普通双相磷酸钙陶瓷(Biphasic calciam phosphate,BCP)作为对照,于4、12、24周取材,测定材料降解率(失重率),从大体、组织学观察以了解材料降解情况。结果:材料肌内植入后降解率测定结果:NanoBCP降解率为32%,BCP的降解率为13%,HA的降解率为3%。组织学观察发现,NanoBCP肌内植入24周后,大部分NanoBCP支架已经将解,并且将解的碎片已埋入纤维结缔组织里。结论:NanoBCP与BCP、HA相比有良好的降解性能。     

The biosynthetic genes for clavulanic acid and cephamycin production occur as a 'super-cluster' in three Streptomyces

Abstract The cosmid cloning vector pHC79 has been used to clone fragments of chromosomal DNA from the Streptomyces: S. clavuligerus, S. jumonjinensis and S. katsurahamanus . These strains all produce both the β-lactam antibiotic, cephamycin and the β-lactamase inhibitor, clavulanic acid. Although structurally related these two β-lactams are known to be derived from different biosynthetic precursors. Hybridisation studies and restriction mapping have shown that the gene clusters encoding the two biosynthetic pathways are chromosomally adjacent in these strains, thus creating a 'super-cluster' of genes involved in both the production and enhancement of activity of a β-lactam antibiotic.