丁酸梭杆菌发酵甘油制备1,3-丙二醇的研究

本研究采用丁酸梭芽孢杆菌(Clostridium butyricum)从甘油发酵制备1,3—丙二醇。该发酵需厌氧培养,发酵培养基为：甘油6％,葡萄糖1％,玉米浆2％,(NH4)2SO40．2％。发酵温度为34℃,pH为6．5～7。在最佳条件下,发酵50h可产1,3—丙二醇40．7g／L,甘油摩尔转化率达68％。     

丁酸梭菌在医学领域中的研究进展

肠道益生菌是定植于人体肠道内,能产生确切健康功效从而改善宿主微生态平衡,对宿主发挥有益作用的一类活性微生物总称。最近越来越多的研究表明肥胖、糖尿病、肠易激综合征、骨质疏松等与肠道菌群密切相关,而丁酸梭菌由于能够促进其他有益菌（乳杆菌、双歧杆菌）生长,具有其他有益生菌没有的特性,受到越来越多研究者的重视。本文通过综述近几年有关丁酸梭菌与神经、内分泌、消化等系统中部分疾病的关系,总结了丁酸梭菌可能具有的治疗机制,并将其与益生菌在治疗骨质疏松症时所发挥的机制进行对比,以展望丁酸梭菌可能具有治疗骨质疏松的潜力。     

Bioconversion of 2-amino acids to 2-hydroxy acids by Clostridium butyricum

Analysis by gas chromatography-mass spectrometry (GC-MS) of 24-h cultures of Clostridium butyricum type strain in synthetic BMG medium supplemented with various 2-amino acids (10 mM) revealed the presence of the corresponding 2-hydroxy acids. C. butyricum was able to bioconvert l-valine, dl-norvaline, l-leucine, dl-norleucine, l-methionine and l-phenylalanine as well as unusual 2-amino acids, i.e., l-2-aminobutyric acid, l-2-amino-4-pentenoic acid, dl-2-aminooctanoic acid, and dl-2-amino-4-phenylbutanoic acid. l-Isoleucine and cycloleucine were not converted into their corresponding 2-hydroxy acids. The bioconversion rate was maximal with dl-norvaline (6.2%). Chiral GC analysis demonstrated that only d-2-hydroxy-4-methylpentanoic acid is formed from l-leucine, indicating that the bioconversion is stereospecific, with inversion of configuration. d-Leucine and d-methionine were also converted to the corresponding 2-hydroxy acids. This observation opens new aspects in the study of C. butyricum and raises questions about the amino acid metabolism by this species.     

Formation of 2-hydroxy-4-methylpentanoic acid from l-leucine by Clostridium butyricum

Abstract Five Clostridium butyricum strains of different origin were grown in trypticase-yeast extract-hemin medium with or without d-glucose (TGYH or TYH medium, respectively) and in a synthetic basal medium with d-glucose (BMG medium). 2-Hydroxy-4-methylpentanoic acid was detected by gas chromatography-mass spectrometry (GC-MS) for the five strains whether grown in TGYH or TYH medium (270 or 170 μM, respectively). In BMG medium supplemented with l-leucine (10 mM), the concentration of this metabolite was strongly increased (2.8 mM versus 10 μM in the control). After culture in TGYH or TYH medium supplemented with l-( methyl -²H₃)leucine, 2-hydroxy-4-([²H₃]methyl)pentanoic acid was detected by GC-MS. This observation demonstrates that C. butyricum is able to convert l-leucine into the corresponding 2-hydroxy acid and opens a new aspect in the study of C. butyricum metabolism.     

酪酸梭菌活菌散在治疗母乳性黄疸中的应用

目的观察在常规治疗的基础上加用酪酸梭菌活菌散（商品名：宝乐安）治疗新生儿母乳性黄疸的疗效。方法176例母乳性黄疸患儿随机分为治疗组和对照组,治疗组90例,对照组86例,对照组采用常规治疗,治疗组在常规治疗基础上加服酪酸梭菌活菌散,2组均不停止母乳喂养,并观察2组患儿日均总胆红素水平及黄疸消退时间的变化。结果治疗组日均胆红素下降值为（58．61±26．52）μmol／L,显著高于对照组（39．12±25．41）μmol／L（P〈0．01）;黄疸消退时间,治疗组为（4．25±2．68）d,显著短于对照组（6．42±2．74）d（P〈0．01）。结论在常规治疗的基础上加用酪酸梭菌活菌散治疗母乳性黄疸,可迅速降低胆红素水平,缩短治疗时间。     

羊源丁酸梭菌HDRyYB1发酵工艺的优化

【目的】目前,国内外鲜有关于羊源丁酸梭菌的报道。本课题选用羊源丁酸梭菌HDRy YB1为研究对象,对其发酵工艺进行优化,为该菌株作为饲料添加剂应用于畜牧业生产奠定基础。【方法】采用Plackett-Burman(PB)试验设计法和响应面法分析并优化显著影响HDRy YB1菌株发酵液中芽胞数的培养基成分。【结果】发酵培养基中的面粉浓度、鱼粉浓度和米粉浓度显著影响发酵液中的芽胞数,优化后的发酵培养基组分(质量体积比)为:面粉3.72%、鱼粉0.90%、米粉3.96%、酵母粉0.60%、Na Cl 0.19%、Mg SO4·7H2O 0.19%、KH2PO4 0.01%、Na HCO3 0.01%、Ca CO3 0.48%;培养参数为:37°C,初始p H为7.2-7.4,瓶装量100/250,接种量3%。在此条件下,HDRy YB1菌株发酵完全(18 h)的芽胞数为1.478×108 CFU/m L,是优化前的2.7倍。【结论】HDRy YB1菌株发酵培养基得到了优化,优化后的培养基可用于后期的扩大发酵试验,验证其在实践生产中的应用价值。     

酪酸梭菌活菌散在迟发型母乳性黄疸中的应用

目的探讨酪酸梭菌活菌散促进迟发型母乳性黄疸消退的疗效。方法将87例迟发型母乳性黄疽患儿随机分为对照组和观察组,对照组43例和观察组44例,对照组采用常规治疗,观察组在常规治疗的基础上加服酿酸梭菌活菌散,500mg/次,3次/d,两组均不停止母乳喂养。观察两组患儿日均胆红素水平的变化及黄疽消退时间。结果观察组日均胆红素下降程度显著大于对照组（P〈0. 01）,观察组黄疸消退时间明显短于对照组（P〈0. 01）,差异有统计学意义。结论在常规治疗的基础上加用酪酸梭菌活菌散治疗迟发型母乳性黄疸,不仅可加速胆红素降低水平,缩短黄疽持续时间,而且能显著缓解茵扼黄颗粒引起的腹泻,是一种简单、安全、有效的治疗措施,并减少停喂母乳对乳母及乳儿的不利影响,值得临床推广应用。     

产1,3-丙二醇菌株丁酸梭菌的诱变育种

甘油由丁酸梭菌转化成1,3-丙二醇的研究是厌氧条件下进行。为了获得1,3-丙二醇的高产突变株,以丁酸梭菌为出发菌株进行诱变处理。经过硫酸二乙酯（DES）化学诱变得到2株高产正突变株C.but2031和C.but2046,再经过紫外线和亚硝基胍（NTG）复合诱变得到突变株C.but3037。经过初筛、复筛和传代实验,表明其是稳定的突变株。C.but3037的1,3-丙二醇产量由出发菌株的2.2g/L提高到15.7g/L,提高了6.13倍,     

Effect of growth supplements and whey pretreatment on butyric acid production by Clostridium butyricum

Improved production of butyrate (up to 19 g/l) from whey by Clostridium butyricum was achieved by adding either yeast extract (5 g/l) or biotin (50 g/l). Hydrolysed lactose and proteolysed whey were less effective even with added biotin.The authors are with the Department of Biochemical Technology, Faculty of Chemical Technology, Slovak Technical University, Radlinského 9, Bratislava 812 37, Slovakia     

仪器法测定口服酪酸梭菌散剂活菌数的研究与应用

目的 利用BD FACSMicroCountTM全自动微生物计数分析系统测定口服酪酸梭菌活菌散剂中的活菌数,以探索BD FACSMicroCountTM全自动微生物计数分析系统测定微生态制剂活菌数的可行性。方法 按照《BD FACSMicroCountTM全自动微生物计数分析系统使用说明书》对该药品的活菌数进行测定。结果 仪器所测得数据与药品包装上理论活菌数基本相符。结论 可使用BD FACSMicroCountTM全自动微生物计数分析系统测定微生态制剂活菌数。     

酪酸梭菌活菌胶囊治疗慢性末端回肠炎疗效评价

目的观察和评价酪酸梭菌活菌胶囊(商品名:阿泰宁)治疗慢性末端回肠炎的临床疗效。方法采用随机对照研究方法,将入组患者随机分为酪酸梭菌联合甲硝唑组、酪酸梭菌组和甲硝唑组,进行治疗,疗程均为4周。治疗期间观察患者的症状、体征及肠镜下变化,治疗结束后评价药物的疗效。结果酪酸梭菌联合甲硝唑组和酪酸梭菌组总有效率分别为97.5%(39/40)和95%(19/20),且治疗中未见不良反应或副作用。酪酸梭菌联合甲硝唑组和酪酸梭菌组总有效率都高于单用甲硝唑组(P<0.01),酪酸梭菌联合甲硝唑组和酪酸梭菌组之间差异无统计学意义(P>0.05)。结论酪酸梭菌活菌胶囊是治疗慢性末端回肠炎的一种安全有效的微生态药品,其疗效优于单用甲硝唑。     

Plasmid localization of a type E botulinal neurotoxin gene homologue in toxigenic Clostridium butyricum strains, and absence of this gene in non-toxigenic C. butyricum strains

It has been shown recently that two Clostridium butyricum strains (ATCC 43181 and ATCC 43755) contain a botulinal neurotoxin type E (BoNT/E) gene closely related to that of C. botulinum type E. In this study, we show that this gene is located on a large plasmid in the two toxigenic C. butyricum strains and is absent in 18 non-toxigenic C. butyricum and C. beijerinckii strains. Interestingly, the 230 bp upstream and the 1260 bp downstream of the neurotoxin coding sequence are not present in either the non-toxigenic C. butyricum or C. beijerinckii strains. Our data suggest a BoNT/E gene transfer from C. botulinum E to originally non-toxigenic C. butyricum strains.     

酪酸菌对动物肠道致病菌体外拮抗作用的研究

探讨了酪酸菌对三种畜禽肠道致病菌的体外拮抗作用。将酪酸菌分别和猪大肠杆菌、鸡大肠杆菌、鸡白痢沙门氏菌三种畜禽肠道致病菌按不同的比例混合接种于CL液体培养基中进行厌氧培养,培养过程中三种致病菌的活菌数急剧下降,30h后的菌落数都逐渐降为零。说明酪酸菌在体外实验中对上述三种畜禽肠道致病菌都有较强的拮抗作用,其中酪酸菌对猪大肠杆菌、鸡大肠杆菌和鸡白痢沙门氏菌的最佳接种比例分别为：10：1～100：1、10：1～100：1和5：1～10：1。     

酪酸梭菌治疗肠易激综合征疗效的Meta分析

目的 系统评价酪酸梭菌治疗肠易激综合征（irritable bowel syndrome,IBS）的临床疗效。方法 计算机检索PubMed、Embase、the Cochrane Library、维普、万方、中国知网和中国生物医学文献数据库,收集建库至2019年5月1日前国内外公开发表的关于酪酸梭菌治疗IBS的随机对照试验。由两位研究员独立进行筛选、资料提取及质量评价,采用RevMan 5.3软件进行Meta分析。结果 最终纳入19篇随机对照试验,共2 194例患者。Meta分析结果显示：酪酸梭菌辅助治疗IBS时,联合治疗组的总有效率明显高于对照组（93.4% vs 77.1%,OR=4.32,95% CI 3.09‒6.04,P<0.05）;而单用酪酸梭菌治疗IBS疗效并不优于常规药物（78.8% vs 68.9%,OR=2.05,95% CI 0.96‒4.38,P=0.06）;且联合用药短期复发率显著降低（14.6% vs 29.2%,OR=0.41,95% CI 0.20‒0.85,P=0.02）。纳入的所有研究均无严重不良反应发生。结论 单用酪酸梭菌与常规药物治疗IBS相比,酪酸梭菌并未显示出更好的疗效。而采用口服酪酸梭菌辅助治疗IBS可显著提高患者的临床疗效、降低复发率,且安全性好。     

酪酸梭菌的研究与应用进展

酪酸梭菌属于微生态制剂,酪酸梭菌具有抑制肠出血性大肠埃希菌、痢疾志贺菌、霍乱沙门菌、霍乱弧菌等肠道致病菌的生长繁殖,抑制艰难梭菌生长繁殖,促进双歧杆菌、乳酸菌等肠道有益菌的生长繁殖,加强肠道黏膜膜营养代谢、保护受损的黏膜屏障,激活免疫细胞、改善并恢复肠道正常免疫力,防治肿瘤的生长的作用.所以酪酸梭菌制剂对腹泻、肠炎、肠易激综合征、结直肠癌新生儿黄疸等疾病都有很强的防治作用.     

酪酸梭菌活菌散辅助治疗新生儿黄疸的随机多中心临床疗效观察

目的观察酪酸梭菌活菌散(商品名:宝乐安)辅助治疗新生儿黄疸的临床疗效。方法将543例新生儿黄疸患儿随机分为观察组和对照组,观察组274例,对照组269例。观察组在蓝光治疗的同时给予酪酸梭菌活菌散口服,0.5g/次,3次/d,服至黄疸消退;对照组只给予蓝光治疗。于治疗后48h、72h、96h检测血清胆红素值。结果治疗72h、96h后观察组血清胆红素水平较对照组明显下降(P0.05)。结论在蓝光治疗新生儿黄疸的同时服用酪酸梭菌活菌散,可迅速降低胆红素水平,缩短治疗时间。     

In vitro evaluation on adherence and antimicrobial properties of a candidate probiotic Clostridium butyricum CB2 for farmed fish

Aims: To characterize the antimicrobial and adhesion ability of candidate probiotic Clostridium butyricum CB2 for farmed fish in vitro. Methods and Results: The potential probiotic Cl. butyricum CB2 had been evaluated for its adhesion capacity and antagonistic effect against two fish pathogens Aeromonas hydrophila and Vibrio anguillarum by the intestinal cell model. In addition, the aggregation ability and antimicrobial property on agar plate were assayed. The results indicated that the candidate probiotic Cl. butyricum CB2 have strong adhesion property and a higher antagonistic activity to Aer. hydrophila and V. anguillarum both on agar plate and cell model. Clostridium butyricum showed a higher aggregation which might be the reasons for bacteria adhesion and antimicrobial activity. Conclusions: The strain Cl. butyricum CB2 could be used as potential probiotic to inhibit pathogens growth and prevent their colonization in fish intestinal tract. Significance and Impact of the Study: This study revealed the antimicrobial and adhesion characteristic of Cl. butyricum CB2 which was selected as the potential probiotic to farmed fish.     

酪酸梭菌活菌散联合抗菌药治疗小儿急性细菌感染性腹泻临床疗效观察

目的观察和评价酪酸梭菌活菌散（商品名：宝乐安）联合抗菌药治疗小儿急性细菌感染性腹泻临床疗效。方法将60例急性细菌感染性腹泻患儿随机分为观察组和对照组,观察组30例,对照组30例,2组均给予口服抗菌药及对症支持治疗。其中观察组在服用抗菌药2h后服用酪酸梭菌活菌散,0.5g/次,3次/d;对照组只给予抗菌药及对症支持治疗,不用酪酸梭菌活菌散。对2组总有效率及体温、大便恢复正常时间进行统计分析。结果观察组总有效率为96.7%,对照组总有效率为83.3%,2组相比差异具有非常显著性（P〈0.01）;观察组体温、大便恢复正常时间均显著短于对照组（P〈0.01）。结论酪酸梭菌活菌散联合抗菌药治疗小儿急性细菌感染性腹泻的疗效,优于单用抗菌药,值得临床推广应用。     

酪酸梭状芽孢杆菌低成本培养基的优化

本文重点在于提高酪酸梭状芽孢杆菌活菌数量,降低发酵培养基成本。通过对发酵培养基中不同碳源、氮源、生长因子等进行单因素研究,得到最佳培养基组成：可溶性淀粉10/L,豆粕（中性蛋白酶水解3h）20g／L,玉米浆3g／L。用此培养基在37℃培养24h,采用高层半固体琼脂试管法对酪酸梭状芽孢杆菌进行活菌计数,活菌数可达8．2×10^8cfu／mL.培养基中添加K2HPO45g／L、MgSO4·7H2O0．2g／L、MnSO3·H2O0．2g/L培养32h时,酪酸梭状芽孢杆菌芽孢转化率可达95％。     

酪酸梭菌活菌散在防治早产儿喂养不耐受方面的应用

目的观察和评价酪酸梭菌活菌散防治早产儿喂养不耐受的临床疗效。方法将56例早产儿随机分成观察组和对照组,观察组30例,对照组26例。对照组给予早产儿配方奶及部分肠外营养等常规治疗,观察组在常规治疗基础上同时添加酪酸梭菌活菌散。观察2组早产儿恢复出生体重、达到全胃肠喂养时间及发生喂养不耐受等情况。结果观察组早产儿恢复出生体重、达到全胃肠喂养时间明显短于对照组(P<0.01或P<0.05),喂养不耐受的发生率也显著少于对照组(P<0.05)。结论早产儿服用酪酸梭菌活菌散,对防治喂养不耐受具有积极的作用,能减少早产儿喂养过程中呕吐、胃潴留、腹胀的发生,促进患儿早期的生长发育,缩短达到全胃肠喂养的时间。