基于转录组学分析揭示蜡样芽胞杆菌(Bacillus cereus)AR1002阻控黄曲霉生长的生理机制研究

黄曲霉(Aspergillus flavus)极易侵染油料等农产品,其次级代谢产物黄曲霉毒素严重危害人体健康,阻控黄曲霉污染已成为亟待解决的国际难题。为探明蜡样芽胞杆菌(Bacillus cereus)AR1002抑制黄曲霉生长、阻控黄曲霉污染的生理及分子机制。该研究采用AR1002代谢产物对黄曲霉进行处理,并对黄曲霉生长及产孢表型进行鉴定,通过显微结构及转录组学分析,对AR1002的抑菌机制进行了初探。蜡样芽胞杆菌AR1002代谢物可抑制黄曲霉菌丝生长、干物质积累分别达24.73%,65.00%,减少孢子数量达98.80%;AR1002通过抑制glpA和AYR1等关键基因表达调控黄曲霉甘油磷酸代谢,通过抑制SEC61A、RAD23、ATP13A1、UBE2G2等关键基因的表达调控内质网中蛋白质加工;此外,AR1002代谢产物通过下调RAD51和RAD54B等DNA修复机制相关基因,抑制黄曲霉同源重组及非同源性末端连接过程,最终达到抑制黄曲霉生长及产孢的目的。上述结果表明,蜡样芽胞杆菌(Bacillus cereus)AR1002有望应用于实际生产中以应对黄曲霉污染问题。

南瓜白粉病拮抗内生细菌Bacillus cereus B4-2的分离鉴定及生防效果

从高抗白粉病的南瓜植株内分离筛选出对南瓜白粉病有较好防治效果的内生拮抗细菌菌株,为南瓜白粉病的生物防治提供新途径。采用孢子萌发抑制法、叶盘漂浮法和温室盆栽试验筛选出有较好防效的内生细菌,用形态观察、生理生化测定和分子生物学方法进行鉴定。结果表明,从高抗白粉病的南瓜自交系22G3植株内分离获得29株内生拮抗细菌菌株,5个菌株对南瓜白粉菌的孢子萌发抑制率在50%以上,其中菌株B4-2对白粉菌的孢子萌发抑制率最高,为88.65%。叶盘和室内盆栽试验结果均表明,菌株B4-2对南瓜白粉病的防治效果最好,防治效果分别达79.58%和83.21%。不同稀释倍数的B4-2发酵液对南瓜白粉病均有一定的防治效果,随着稀释倍数的增大,防治效果降低,其中菌株B4-2发酵液原液对南瓜白粉病的防治效果最好,达82.14%。通过形态特征、生理生化特征及基因序列分析,鉴定菌株B4-2为蜡样芽孢杆菌(Bacillus cereus)。综上所述,蜡样芽孢杆菌B4-2能较好地防治南瓜白粉病,有一定的开发潜力。

蚯蚓联合生防细菌Bacillus velezensis改善连作百合土壤细菌群落结构及防治枯萎病的效果

【目的】从土壤理化性状和微生物群落结构及多样性角度,研究蚯蚓联合生防细菌改善百合连作障碍的机理和效果,为湖南百合产业化种植提供技术措施。【方法】采用盆栽试验方法,在湖南长沙连续种植了两季百合(Lilium lancifolium Thunb.)。供试土壤为红壤,生防菌为Bacillales velezensis YFB3-1菌液(菌体浓度为10^(9) CFU/mL),蚯蚓为赤子爱胜蚓(Eisenia fetida)。试验设单施菌液(T1)、只接种蚯蚓(T2)、同时使用菌液和蚯蚓(T3) 3个处理,以不使用菌剂和蚯蚓为对照(CK)。调查了两茬百合产量和枯萎病病情指数,分析了第二茬百合根际土壤理化性质、细菌群落结构和多样性,以及连作百合产量与根际土壤理化性质、细菌群落和枯萎病的相关性。【结果】与第一茬相比,同一处理的第二茬百合产量都显著下降。两年连作百合产量都以同时使用菌液和蚯蚓(T3)处理为最高,分别为16464、15674 kg/hm^(2),都显著高于同年份其他处理。两年连作百合枯萎病发病率和病情指数都以T3为最低,都显著低于同年份其他处理,T3处理百合枯萎病病情指数分别为18.15(2019年)、25.00 (2020年),并且两年的防治效果(T3)都明显优于单施菌液(T1)和接种蚯蚓(T2)。相比于对照(CK),2019、2020年T3处理百合枯萎病防治效果分别为44.55%、37.66%,分别增产15.05%、14.78%。蚯蚓联合生防菌YFB3-1增加了根际土壤Rhizobiales、Flavobacteriales、Pseudomonadales、Bacillales等有益微生物的种群丰度,蚯蚓与生防菌YFB3-1在抑制连作百合枯萎病方面具有显著的互作效应,因而缓解连作百合产量下降的效果最佳。【结论】在施用芽孢杆菌生防菌剂防治连作百合枯萎病的同时,可以增施有机肥(牛粪)来提高土壤中蚯蚓数量,有效预防百合枯萎病发生,减少连作百合产量损失。

莫海威芽孢杆菌(Bacillus mojavensis)ZLX10对发酵雪茄烟叶中性致香成分的影响

为探究烟草源微生物单菌发酵对雪茄烟叶挥发性致香成分的影响,采用GC-MS检测莫海威芽孢杆菌(Bacillus mojavensis)ZLX10发酵雪茄烟叶中性致香成分,筛选差异致香成分,并与感官评价进行相关性分析,结合ROAV值变化,明确ZLX10提升雪茄烟叶感官品质的关键致香成分。结果表明,ZLX10发酵雪茄烟叶后,香气总量和大多数中性致香成分含量均显著提高,共筛选得到15种差异中性致香成分,共有10种致香成分显著上调(P<0.05),其中巨豆三烯酮A、巨豆三烯酮B、6-甲基-5-庚烯-2-酮、2,6,6-三甲基-2-环己烯-1,4-二酮、茄酮和植物醇与烟叶香气呈显著正相关(P<0.05),可认为是增加烟叶干香和花香的关键差异致香成分;共有5种致香成分显著下调(P<0.05),其中苯乙醛和2,4-二叔丁基-6-硝基苯酚与烟叶刺激性呈显著正相关(P<0.05),对烟叶香气有较大的负面影响。由此可知,ZLX10发酵能促进雪茄烟叶香气质和香气量得到提升,这为人工接菌发酵雪茄烟叶提供了基础。

1株抗真菌贝莱斯芽孢杆菌(Bacillus velezensis)lut-Y1的鉴定及抑菌活性研究

【目的】探究1株来自枸杞果实的贝莱斯芽孢杆菌lut-Y1的生长特性及抑菌活性,为微生态制剂的开发和应用提供参考。【方法】通过形态观察和16S rDNA基因测序分析对该菌株进行鉴定;测定该菌株生长曲线及最适pH、碳源、氮源及盐耐受度;采用平板对峙法观察该菌株对链格孢菌(Alternaria alternate)、小麦根腐离蠕孢(Bipolaris sorokiniana)和小孢壳二孢(Ascochyta leptospora)的颉颃作用,并分析该菌株发酵液对3种病原真菌的抑菌活性。【结果】lut-Y1菌株在LB、NA和PDA培养基上均生长良好。根据形态和16S rDNA基因序列特征将lut-Y1菌株鉴定为贝莱斯芽孢杆菌(Bacillus velezensis)。在LB液体培养基中,lut-Y1菌株在0~3 h为迟滞期,3~18 h为对数期,18~62 h为稳定期,62 h后进入衰亡期;最适pH为7.0,最适碳源和氮源分别为蔗糖和蛋白胨,对NaCl的耐受量不超过2.0 g/L。lut-Y1菌株对链格孢菌有明显的颉颃作用,对小麦根腐离蠕孢和小孢壳二孢的颉颃作用较弱。对峙平板的两菌交界处,链格孢菌基内菌丝细弱、弯曲;小麦根腐离蠕孢基内菌丝细胞壁被破坏,有泡状突起。发酵液添加量为25%时生长圈直径分别缩小72.80%和83.24%。【结论】贝莱斯芽孢杆菌lut-Y1具有较好的抑菌活性,可为开发以芽孢杆菌为主要活性成分的微生态制剂提供材料。

不同有机废弃物载体对Bacillus megaterium有效性的影响

为了给Bacillus megaterium选择理想的吸附载体。测定了草炭、金针菇渣、腐熟鸡粪、牛粪、羊粪的吸菌量及其载体菌剂的有效活菌数、胞外多糖(EPS)含量和发芽指数。草炭的吸菌量最高为1.64 mL/g,但随存放时间延长,草炭载体菌剂的有效活菌数和EPS含量急剧降低,且38 d内的发芽指数均小于80%,表现植物毒性。鸡粪和金针菇渣的吸菌量虽小于草炭,但随存放时间延长,鸡粪载体菌剂的有效活菌数和EPS含量逐渐增大,金针菇渣载体菌剂的EPS含量逐渐增加、有效活菌数降低幅度缓慢,且存放7 d后发芽指数均大于80%,表现植物无毒性。第38天时鸡粪和金针菇渣载体菌剂的有效活菌数分别为1.29×10^(9)CFU/g和6.20×10^(8)CFU/g,EPS含量分别为535.29 mg/kg和559.34 mg/kg,均显著高于其他载体菌剂。鸡粪和金针菇渣是B.megaterium较为理想的吸附载体。

Mn^(2+)促进Bacillus altitudinis LZP02生物膜形成

【目的】Bacillus altitudinis LZP02是一株水稻根际促生菌(PGPR),能定殖于水稻根部,形成生物膜,且Mn^(2+)对LZP02菌株生物膜形成具有促进作用,但调控机制尚不明确,旨在探究Mn^(2+)对LZP02菌株生物膜形成的促进机制。【方法】采用结晶紫染色法进行生物膜定量分析、蒽酮-硫酸法测定胞外多糖产量、扫描电镜(SEM)观察LZP02菌株在水稻根际定殖情况、转录组学测序技术分析差异表达基因(DEGs)。【结果】添加4 mmol/L Mn^(2+)和8 mmol/L Mn^(2+)显著提高了LZP02菌株的成膜能力和胞外多糖产量,扫描电镜发现4 mmol/L Mn^(2+)和8 mmol/L Mn^(2+)提高了LZP02菌株在水稻根际的定殖能力,随着Mn^(2+)浓度的增加,DEGs数量显著增多,其主要富集在芽孢形成、毒素代谢途径和双组分系统中。1 mmol/L Mn^(2+)和4 mmol/L Mn^(2+)处理组发现skf操纵子中基因的表达均上调。扫描电镜观察发现在1 mmol/L Mn^(2+)和4 mmol/L Mn^(2+)处理组中LZP02菌体存在损伤。4 mmol/L Mn^(2+)处理组双组分系统中KinE和Spo0A基因均显著上调。【结论】Mn^(2+)通过“嗜食同类”和双组分系统中KinE基因激活Spo0A~P提高了菌株LZP02的成膜能力。

纳豆芽孢杆菌Bacillus natto NK4液态发酵产纳豆激酶的工艺优化

为提高纳豆激酶的产量,采用单因素试验及正交试验对纳豆芽孢杆菌Bacillus natto NK4进行发酵条件优化,获得该菌种液体发酵产酶的最优条件。结果表明,纳豆芽孢杆菌产纳豆激酶最佳的发酵条件为接种量2%,培养基初始pH值6.0,培养温度34℃,发酵时间72 h。此时,纳豆激酶酶活力由848.28 IU/mL提高到1569.31 IU/mL。

生物炭载Bacillus subtilis菌粒对连作芹菜生长的影响

为解决土壤连作障碍对蔬菜生长发育的不利影响,本研究以连作3年的芹菜土壤为研究对象,采用随机区组试验,设置5种处理:单独施加生物炭(BC)、单独施加Bacillus subtilis菌剂(BS)、生物炭与菌剂混合施加(BB)、施加生物炭载B.subtilis菌粒(BP)、不施加生物炭与菌剂(CK),盆栽种植芹菜,测定其生长与品质指标,探究生物炭和B.subtilis对连作芹菜生长的影响。结果表明,BB、BP处理组芹菜的生长和品质指标都高于其他土壤处理组,BP处理组的芹菜鲜质量、可溶性蛋白含量以及纤维素含量均显著高于BB处理组,分别为60.25 g/棵、27.25 mg/g、2.48%,其他指标两处理间无显著差异。不同土壤处理组芹菜的生长和品质指标都高于CK处理,其中BP处理组各指标值最高,尤其是芹菜鲜质量,增幅高达226.56%。因此,施加生物炭载B.subtilis菌粒提高了连作芹菜生长指标与品质,为农业生产上缓解蔬菜连作障碍奠定了理论基础。

Bacillus globisporus N75 6-α-葡萄糖基转移酶在枯草芽孢杆菌中的重组表达

6-α-葡萄糖基转移酶具有催化α-1,4-葡聚糖的外切水解和转糖基化功能,可通过转糖基活性产生非还原端连有α-1,6键的α-葡聚糖,可用于生产环交替糖及其分支衍生物和环异麦芽寡糖等新型功能糖。为了更好地将6-α-葡萄糖基转移酶应用于食品领域,该研究对来源于Bacillus globisporus N75的6-α-葡萄糖基转移酶在食品安全级菌株枯草芽孢杆菌中进行重组表达,并通过表达宿主的选择及优化发酵条件进一步提高重组蛋白产量。结果表明,宿主WHS9更有利于6-α-葡萄糖基转移酶的重组表达,在20℃的发酵条件下,胞内外可溶性总酶活力可达2.11 U/mL;在25℃添加20 mmol/L海藻糖发酵48 h后,总酶活力可达到2.21 U/mL;优化发酵时间后,发酵36 h总酶活力可达3.14 U/mL,是优化前WHS9表达酶活力的196倍。该研究首次实现了6-α-葡萄糖基转移酶在枯草芽孢杆菌中的重组表达。

李氏禾内生细菌Bacillus cereus J01吸附Cr^(3+)的研究

为考察李氏禾内生细菌蜡样芽孢杆菌Bacillus cereus J01菌株吸附Cr^(3+)的性能,以蜡样芽孢杆菌Bacillus cereus J01失活菌体作为生物吸附剂,采用单因素实验方法考察pH、温度、Cr^(3+)初始浓度、吸附剂用量、吸附时间等条件对Cr^(3+)吸附性能的影响,分析其等温吸附过程、动力学过程及热力学过程,并采用红外光谱方法对吸附机理进行初步分析。结果表明:(1)在50 mL反应体系中,当pH值为6、温度为40℃、Cr^(3+)初始浓度为150 mg·L^(-1)、吸附剂投加量为0.2 g、吸附时间为12 h时,蜡样芽孢杆菌Bacillus cereus J01失活菌体对Cr^(3+)的吸附性能达到最佳,其平衡吸附量和Cr^(3+)去除率分别为34.30 mg·g^(-1)、91.60%。(2)等温吸附过程分析结果显示,朗格缪尔吸附等温模型能更好地模拟蜡样芽孢杆菌Bacillus cereus J01失活菌体对Cr^(3+)的吸附,其吸附过程更倾向于单分子层吸附。(3)吸附过程动力学分析结果显示,蜡样芽孢杆菌Bacillus cereus J01失活菌体对Cr^(3+)的吸附更符合准二级动力学速率方程。(4)吸附过程热力学分析结果显示,在40℃下,吸附过程的△G、△H和△S分别为-2.609 kJ·mol^(-1)、61.792 kJ·mol^(-1)和206.11 J·mol^(-1),该温度下吸附过程是自发过程。(5)红外光谱分析结果显示,蜡样芽孢杆菌Bacillus cereus J01失活菌体对Cr^(3+)的吸附可能通过细胞成分中的氨基、羟基和羰基起作用。该研究结果表明该菌的失活菌体对Cr^(3+)具有较强的吸附性能,并在铬污染的环境治理中具有较好的应用潜力。

耐低温降解菌Bacillus mobilis10的生物学特性及其对海参养殖水质的净化作用

以分离自沿海土壤中的1株芽孢杆菌10为试验对象,研究其在低温条件下(15℃)的生物学特性,实现该菌剂的中试工程化生产以及在海参养殖中对水质净化的示范应用。该菌株通过形态观察及16S rRNA基因序列分析被鉴定为Bacillus mobilis。15℃,经透明圈法确定该菌株具有蛋白酶活性、淀粉酶活性和纤维素酶活性;发酵56 h蛋白酶活性达到9.05 U/mL;在8%质量分数NaCl条件下仍可以存活。利用海参饵料培养基检测菌株对溶解物的降解能力,试验结果显示,B.mobilis 10接种8 h后对饵料培养基中蛋白的降解率达(36.72±0.53)%,对化学需氧量的清除率达(22.80±0.81)%。中试规模(50 L)发酵制备微生物制剂并以终密度约1.0×10^(4)cfu/mL的菌液泼洒海参池,对水体中B.mobilis 10的活菌数、pH、蛋白质含量及化学需氧量进行为期8 d的动态监测,结果表明,以B.mobilis 10制备的微生物制剂能够显著降低海参养殖水体中蛋白和化学需氧量,起到净化水质的目的。试验结果可为水产生态养殖及微生物制剂的开发提供理论基础和应用示范实例。

矿化细菌Bacillus paramycoides对重金属Pb(Ⅱ)和Cd(Ⅱ)的去除及矿化特性

微生物诱导生物矿化可有效固定重金属离子,被认为是一种重要的重金属污染环境修复工艺,对重金属污染生态修复有重要的现实意义。从贵州黔西北某铅锌矿区周边土壤中分离出一株耐Pb(Ⅱ)和Cd(Ⅱ)菌株GS-2,经形态学特征和16S rRNA鉴定为Bacillus paramycoides,菌株GS-2对Pb(Ⅱ)和Cd(Ⅱ)的耐受分别高达1500和50 mg/L,对250 mg/L Pb(Ⅱ)和5 mg/L Cd(Ⅱ)去除率分别为99.54%和69.04%,对重金属Pb(Ⅱ)和Cd(Ⅱ)(均为25 mg/L)混合去除率分别为99.73%和71.46%。SEM-EDS、FTIR和XRD结果显示,菌株通过表面的羟基、羧基、酰胺基和磷酸基团等官能团将Pb(Ⅱ)和Cd(Ⅱ)诱导矿化成Ca_(2.5)Pb_(7.5)(OH)_(2)(PO_(4))_(6)、Ca_(2.1)Pb_(7.9)Cl_(2)(PO_(4))_(6)、Pb_(3)(PO_(4))_(2)、Pb_(5)(PO_(4))_(3)OH、Pb_(9)(PO_(4))_(6)、Pb_(10)(PO_(4))_(6)(OH)_(2)和Ca_(0.67)Cd_(0.33)CO_(3)等晶体矿物。菌株GS-2具有较好的去除和矿化能力,对环境中Pb(Ⅱ)和Cd(Ⅱ)污染的生物修复具有一定应用潜力。

Antiseptic Efficacy of A Soap Made from Biosurfactants Isolated from Bacillus and Lactobacillus against Pathogenic Bacteria

The aim of our study was to use a biosurfactant produced by Bacillus and Lactobacillus isolates as an antiseptic in the formulation of local soap. A total of 60 isolates were characterized by microbiological techniques (30 Bacillus and 30 Lactobacillus) and the ability to produce biosurfactants was demonstrated by a hydrocarbon emulsification index (E24). The emulsification indexes (E24) varied from 9% to 100% for Bacillus and from 33% to 100% for Lactobacillus as well. The antagonistic assay showed that biosurfactants were able to inhibit the formation of biofilms and growth of pathogens such as Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus cereus, Salmonella typhirium, Shigella boydii and Proteus mirabilis. The biosurfactant consortium (BioC) from Bacillus consortium and from Lactobacillus was able to inhibit biofilm formation and the pathogens growth. The BioC was stable to alkaline pH and the temperatures stability of Biosurfactant was ranging from 50°C to 90°C. The soap was made by the cold saponification process using one biosurfactant consortium formulated. This soap has a pH of 10 and showed good cleaning power and good foam stability. Similarly, the soap showed good antiseptic power and disinfection power against all pathogens tested. Handwashing is critical to preventing disease transmission. The persistence of pathogens in waste water was evaluated. The BioS produced showed good disinfection power against all pathogens tested. The valor of reduction on the hands and in the waste water was significantly more than compared to the control soaps used. This soap could be used in the prevention, fighting, and treatment of bacterial and viral infections.

Bacillus tropicus来源β-D-呋喃果糖苷酶基因的克隆与表达及其酶学特性研究

为获得新β-D-呋喃果糖苷酶资源并探究其酶学性质及在转化糖中的应用潜力,挖掘出Bacillus tropicus来源的β-D-呋喃果糖苷酶基因BtFFase8,成功合成后表达在大肠杆菌宿主中。通过数据库筛选β-D-呋喃果糖苷酶的基因序列,将基因克隆至载体pET-28a(+)中。将获得的重组质粒转化至大肠杆菌BL21,构建重组菌株大肠杆菌BL21/pET28a-BtFFase8。采用亲和层析纯化克隆酶,用SDS-PAGE法分析蛋白分子量,通过葡萄糖试剂盒检测酶水解产物的能力得到酶活力,用福林酚试剂法测定蛋白质含量。克隆酶BtFFase8的分子质量为57.0 kDa,对蔗糖底物的比酶活力为13.4 U/mg;BtFFase8的最适pH和最适温度分别为pH 6.5和45℃,且在pH 5.0~8.0和40℃及以下温度保持酶活力稳定,残余酶活力大于80%;BtFFase8能耐受碱性蛋白酶、酸性蛋白酶和胰蛋白酶的水解,残余酶活力大于70%。BtFFase8的发现及克隆酶优良的酶学功能为其未来应用于食品加工,特别是在转化糖的生产中奠定了良好的理论基础。

Research advance of Bacillus velezensis:bioinformatics,characteristics,and applications

Bacillus velezensis is a Gram-positive and spore-forming bacterium.It has potent antimicrobial properties that can be used to promote plant growth and as a pesticide by inhibiting pathogens.B.velezensis has the capability to generate a diverse range of enzymes that have potential applications in various fields,such as enzyme production,fermented food,degradation of pollutants,and bioenergy.In addition,B.velezensis is a promising probiotic.It possesses high bile-salt tolerance characteristics and has a high success rate of colonization in the intestinal mucosa.Besides,the strain can also regulate gut microbiota constitute by increasing the number of beneficial microorganisms and decreasing the number of pathogens.Furthermore,based on its special properties,including high-yield protease production and high salt-tolerance,B.velezensis shows potential for use in marine protein fermentation,opening up new avenues for the development of novel food products and bioactive peptides.In addition,B.velezensis can shorten the fermentation time as well as improve the nutritional value and flavor of fermented food.The safety of B.velezensis for food production was evaluated.This review provides valuable insights into the potential uses and benefits of B.velezensis,particularly in the context of fermented foods.

Induced CTL-S15 gene expression by Bacillus thuringiensis declines susceptibility in Spodoptera exigua

It has been reported that C-type lectins(CTLs),which are pattern recognition receptors of the insect innate immunity response,may compete with Cry toxin for the receptor alkaline phosphatase to decrease its toxicity in insects.However,to date,which CTLs affect larval susceptibility to Bt in Spodoptera exigua is not clear.In this study,33 CTL genes were identified from S.exigua.Based on the number of carbohydrate-recognition domains(CRDs)and the domain architectures,they were classified into three groups:(1)nineteen CTL-S(single-CRD),(2)eight immulectin(dual-CRD)and(3)six CTL-X(CRD with other domains).RT-qPCR analysis revealed that expression levels of SeCTL-S15,IML-4 and CTL-X6 were upregulated after challenge with Bt and Cry1Ab.Tissue and developmental stage expression analysis showed that only SeCTL-S15 was mainly expressed in the midgut and larva,respectively.Knockdown of SeCTL-S15 significantly increased Bt susceptibility,as indicated by reduced survival and larval weight.These results suggest that CTL-S15 might play a vital role in the low susceptibility of larvae to Bt in S.exigua.Our results provide new insights into CTL function in insects.

Enhanced extracellular production of alpha-lactalbumin from Bacillus subtilis through signal peptide and promoter screening

Alpha-lactalbumin(α-LA)is a major whey protein found in breast milk and plays a crucial role in the growth and development of infants.In this study,Bacillus subtilis RIK1285 harboring AprE signal peptide(SP)was selected as the original strain for the production ofα-LA.It was found thatα-LA was identified in the pellet after ultrasonic disruption and centrifugation instead of in the fermentation supernatant.The original strain most likely only producedα-LA intracellular,but not extracellular.To improve the expression and secretion ofα-LA in RIK1285,a library of 173 homologous SPs from the B.subtilis 168 genome was fused with target LALBA gene in the pBE-S vector and expressed extracellularly in RIK1285.SP YjcN was determined to be the best signal peptide.Bands in supernatant were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and purified by nickel column to calculate the highest yield signal peptide.In addition,different promoters(P_(aprE),P_(43),and P_(glv))were compared and applied.The results indicated that the strain RIK1285-pBE-P_(glv)-YjcN-LALBA had the highestα-LA yield,reaching 122.04μg/mL.This study demonstrates successful expression and secretion of humanα-LA in B.subtilis and establishes a foundation for simulating breast milk for infant formulas and developing bioengineered milk.

Bioleaching of vanadium from stone coal vanadium ore by Bacillus mucilaginosus:Influencing factors and mechanism

Vanadium and its derivatives are used in various industries,including steel,metallurgy,pharmaceuticals,and aerospace engineering.Although China has massive reserves of stone coal resources,these resources have low grades.Therefore,the effective extraction and recovery of metallic vanadium from stone coal is an important way to realize the efficient resource utilization of stone coal vanadium ore.Herein,Bacillus mucilaginosus was selected as the leaching strain.The vanadium leaching rate reached 35.5%after 20 d of bioleaching under optimal operating conditions.The cumulative vanadium leaching rate in the contact group reached 35.5%,which was higher than that in the noncontact group(9.3%).The metabolites of B.mucilaginosus,such as oxalic,tartaric,citric,and malic acids,dominated in bioleaching,accounting for 73.8%of the vanadium leaching rate.Interestingly,during leaching,the presence of stone coal stimulated the expression of carbonic anhydrase in bacterial cells,and enzyme activity increased by 1.335-1.905 U.Enzyme activity positively promoted the production of metabolite organic acids,and total organic acid content increased by 39.31 mg·L^(-1),resulting in a reduction of 2.51 in the pH of the leaching system with stone coal.This effect favored the leaching of vanadium from stone coal.Atomic force microscopy illustrated that bacterial leaching exacerbated corrosion on the surface of stone coal beyond 10 nm.Our study provides a clear and promising strategy for exploring the bioleaching mechanism from the perspective of microbial enzyme activity and metabolites.

Oral administration of Bacillus coagulans TQ-35 alleviates allergic responses in OVA-sensitive BALB/c mice

Bacillus coagulans has been extensively studied so far,but there has been a lack of research on its usage in allergy.In this study,we designed to assess the effect of different concentrations of B.coagulans on food allergy in a BALB/c mouse model of ovalbumin(OVA)-induced food allergy and its effect on gut microbes.The assessment of symptoms,specific immunoglobulin E(IgE),T-cell differentiation,and related gene expression levels in sensitized mice by assay indicated that high doses of oral B.coagulans could alleviate allergic symptoms.Treatment with B.coagulans,in the high-dose group,significantly reduced IgE and IgG1 levels and modulated the balance of T helper type 1 cell(Th1)and Th2 and the expression of relevant genes in the spleen.16S rRNA analysis showed that probiotics improved the structure of the microbiota,in particular by boosting the percentage of Clostridia,Bacteroides vulgatus and Enterococcus faecium,and by increasing the abundance of microbial species,thereby modulating the immune system.Therefore,this study can provide insights into the practical application of B.coagulans doses to alleviate OVA allergy.