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Comparative susceptibility and immune responses of Asian and European honey bees to the American foulbrood pathogen, paenibacillus larvae

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Insect Science 2019年第5期26卷 831-842页

作者： Sasiprapa Krongdang Jay D.Evans Yanping Chen Wannapha Mookhploy Panuwan Chantawannakul 1Bee Protection Laboratory Department of BiologyFaculty of ScienceChiang Mai UniversityChiang MaiThailand Interdisciplinary Program in Biotechnology Graduate SchoolChiang Mai UniversityChiang MaiThailand USDA-ARS Bee Research LaboratoryBeltsvilleMarylandUSA Environmental Science Research Center Faculty of ScienceChiang Mai UniversityChiang MaiThailand

American foulbrood (AFB) disease is caused by paenibacillus larvae. Currently, this pathogen is widespread in the European honey bee— Apis mellifera. However, little is known about infectivity and pathogenicity of P. lan'ae in the Asiatic cavity-nesting honey bees, Apis cerana. Moreover, comparative knowledge of P. larvae infectivity and pathogenicity between both honey bee species is scarce. In this study, we examined susceptibility, larval mortality, survival rate and expression of genes encoding antimicrobial peptides (AMPs) including defensin, apidaecin, abaecin, and hymenoptaecin in A. mellifera and A. cerana when infected with P. larvae. Our results showed similar effects of P. larvae on the survival rate and patterns of AMP gene expression in both honey bee species when bee larvae are infected with spores at the median lethal concentration (LC5 0 ) for A. mellifera. All AMPs of infected bee larvae showed significant upregulation compared with noninfected bee larvae in both honey bee species. However, larvae of A. cerana were more susceptible than A. mellifera when the same larval ages and spore concentration of P. larvae were used. It also appears that A. cerana showed higher levels of AMP expression than A. mellifera. This research provides the first evidence of survival rate, LC50 and immune response profiles of Asian honey bees, A. cerana, when infected by P. larvae in comparison with the European honey bee, A. mellifera.

关键词： Apis cerana Apis mellifera honey bee immune gene paenibacillus larvae susceptibility

paenibacillus puldeungensis LK18普鲁兰酶基因的克隆表达及酶学性质研究

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现代食品科技 2019年第7期35卷 107-113,35页

作者：苏红玉崔堂兵华南理工大学生物科学与工程学院

本研究根据类芽孢杆菌的普鲁兰酶基因的保守区设计兼并引物,从paenibacillus puldeungensis LK18中扩增出普鲁兰酶基因(pulA),将其连接至pET-28a(+)载体上,构建出重组表达载体pET-28a(+)-pulA,转入到Escherichia coli BL21(DE3)中,成功地表达了重组普鲁兰酶。结果表明:该普鲁兰酶基因全长1968 bp,编码655个氨基酸。通过Ni柱亲和层析纯化出重组蛋白,测定其比酶活为508.8 U/mg,分子量约为76.95 ku。该重组酶的最适反应温度为45℃,在35℃~40℃下保温120 min后剩余酶活达60%以上;最适作用pH为6.0,在pH 6.0~8.0条件具有较好的稳定性;10 mmol/L的K^+和Mg^2+对该重组普鲁兰酶有激活作用,而Zn^2+、Mn^2+、Ni^2+、Fe^2+、Cu^2+、Co^2+、Ca^2+等对其有不同程度的抑制作用。本研究成功地构建出一株可高效表达普鲁兰酶的重组菌株,具备一定的工业应用价值。

关键词： paenibacillus puldeungensis 普鲁兰酶克隆表达酶学性质

Inhibitory Activity of paenibacillussp. Isolated from Soil in Gotsu City, Shimane Prefecture, Against Xanthomas oryzaepv. oryzae, the Causal Agent of Rice Bacterial Leaf Blight

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Advances in Microbiology 2018年第3期8卷 197-210页

作者： Mohammad Naser Haidary Tomoko Tamura Makoto Ueno Laboratory of Plant Pathology Faculty of Life and Environmental ScienceShimane UniversityMatsueJapan

Microorganism isolates (n = 49) were obtained from the soil samples collected from field in Gotsu city (Kawahira), Shimane. Isolate GT2-E culture inhibited the growth of Xanthomonas oryzae pv. oryzae in disk diffusion method. Rice bacterial leaf blight was suppressed by GT2-E culture in the pre- and post-treated rice leaves. Sequence analysis of 16S rDNA region of the GT2-E isolate indicated that it shared 99% similarity with paenibacillus polymyxa. The growth of GT2-E on LB medium was observed at 15°C, 28°C, 37°C, and 45°C, but not at 4°C. GT2-E isolate could be grown even in the presence of agrochemicals (Amister, Blasin and Kasumin). Furthermore, the growth of X. oryzae pv. oryzae was inhibited by the culture filtrate of GT2-E isolate in disk diffusion method. However, the inhibitory activity of the culture filtrate was heat-unstable. This result suggested that GT2-E isolate can produce heat-unstable inhibitory compound(s). In conclusion, GT2-E isolate might contribute to the development of a new bactericide and biological agent against rice bacterial leaf blight.

关键词： Rice Bacterial Leaf Blight Xanthomonas oryzae pv. oryzae Microorganisms paenibacillus polymyxa

paenibacillus polymyxaZJ-9混合发酵菊粉和葡萄糖合成R,R-2,3-丁二醇

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食品与发酵工业 2015年第3期41卷 8-13页

作者：徐尤勇高健徐虹曹灿黄巍巍南京工业大学食品与轻工学院材料化学工程国家重点实验室江苏南京211816 盐城工学院化学与生物工程学院江苏盐城224051

在7.5 L发酵罐上考察了paenibacillus polymyxa ZJ-9混合发酵菊粉和葡萄糖合成R,R-2,3-丁二醇的工艺条件。选用菊芋菊粉粗提液为发酵前期底物,分析比较不同初始菊粉浓度下的细胞比生长速率(μ)和产物比合成速率(qp),进而研究了补糖种类... 详细信息

在7.5 L发酵罐上考察了paenibacillus polymyxa ZJ-9混合发酵菊粉和葡萄糖合成R,R-2,3-丁二醇的工艺条件。选用菊芋菊粉粗提液为发酵前期底物,分析比较不同初始菊粉浓度下的细胞比生长速率(μ)和产物比合成速率(qp),进而研究了补糖种类和不同补料方式对合成R,R-2,3-丁二醇的影响。结果表明,初始菊粉75.0g/L,当发酵到24 h、31 h时,分别添加15.0 g/L的葡萄糖,发酵效果最佳,44 h时,产物产量达到最高值47.8 g/L,与分批发酵相比,糖转化率由原来的34.9%提高到45.5%,生产强度由原来的0.70 g/(L·h)提高到1.09 g/(L·h),并且副产物乙偶姻、残糖浓度相对较低。

关键词： R,R-2,3-丁二醇 paenibacillus polymyxa ZJ-9 补料分批发酵

paenibacillus bovis BD3526产胞外多糖的培养条件优化

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食品与发酵工业 2016年第5期42卷 108-113页

作者：崔燕丽韩瑨吴正钧光明乳业股份有限公司乳业研究院乳业生物技术国家重点实验室上海2004361 上海海洋大学食品学院上海201306

利用单因素试验和响应面法对paenibacillus bovis BD3526产胞外多糖发酵条件工艺参数进行优化。结果表明:培养温度、时间、培养基装量与paenibacillus bovis BD3526胞外多糖产量存在显著的相关性。当麸皮质量浓度为30 g/L时,BD3526产胞... 详细信息

利用单因素试验和响应面法对paenibacillus bovis BD3526产胞外多糖发酵条件工艺参数进行优化。结果表明:培养温度、时间、培养基装量与paenibacillus bovis BD3526胞外多糖产量存在显著的相关性。当麸皮质量浓度为30 g/L时,BD3526产胞外多糖的优选条件为:培养温度29.91℃、培养时间23.96 h、培养基装量59.58m L/250 m L锥形瓶,采取上述优选条件进行培养,理论预测最大胞外多糖产量为11.11 g/L。验证试验实际测得多糖平均产量为10.92 g/L,比理论预测值低1.71%。

关键词： paenibacillus bovis BD3526 胞外多糖响应面法

Purification and Characterization of Alkaline Xylanase Secreted from paenibacillus macquariensis

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Advances in Microbiology 2013年第1期3卷 32-41页

作者： Meeta Sharma Saurabh Mehta Anil Kumar School of Biotechnology Devi Ahilya University Indore India

An alkaline xylanase secreted by paenibacillus macquariensis RC 1819 has been purified using ammonium sulfate fractionation, ion exchange chromatography using DEAE-cellulose and gel filtration chromatography over Sephadex G-200 and Sephadex G-100. The purified enzyme had the specific activity, 25.2 units/mg protein with birchwood xylan as a substrate. The purified enzyme showed a single protein band over sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight of the enzyme has been found to be 31,000 ± 2000 as determined by using Sephadex G-200 gel filtration chromatography. The subunit molecular weight has also been found to be ~31,000 as determined using SDS-PAGE indicating monomeric enzyme. The enzyme showed optimum activity at pH 8.6 and temperature, 50°C. The Michaelis constant (Km) of the enzyme for birchwood xylan was 2.2 mg/ml as determined using velocity saturation plot. The metal ions viz. Co+2 and Mn+2 stimulated xylanase enzyme activity whereas Hg+2 inhibited the enzyme activity.

关键词： Xylanase paenibacillus macquariensis Purification Michaelis Constant Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis

paenibacillus Infection in a Pediatric Patient on ECMO

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Advances in Infectious Diseases 2022年第1期12卷 74-79页

作者： Wetstein Pace Holloway Adrian Garber Nan Patel Ripal Biology Stevenson University Owings Mills USA Pediatrics Pediatric Critical Care PICU University of Maryland Medical Center Baltimore USA Department of Pediatrics West Virginia School of Medicine Morgantown WV USA

paenibacillus is a spore forming gram positive rod that is usually found in the environment. We describe a case of a patient who contracted this organism having never left the hospital after birth. This neonate contracted paenibacillus septic shock requiring support with venous-arterial extracorporeal membrane oxygenation (ECMO) while still admitted to the hospital after birth. This patient initially presented in severe septic shock due to a Streptococcus agalactiae infection requiring hemodynamic support with ECMO. Following treatment for the Streptococcus agalactiae infection, and while still on ECMO support, the blood culture became positive for paenibacillus. Given that our patient had never left the hospital after birth, the finding of this organism in the blood is unique. The primary defense against this bacterium is usually the skin. The only invasive procedure this patient had was ECMO cannulation which is done in a sterile fashion. Most species are susceptible to vancomycin, clindamycin, fluoroquinolones, aminoglycosides, carbapenems, penicillin, and cephalosporins. This patient was treated with penicillin G for 14 days for the S. agalactiae infection prior to the blood culture being positive for paenibacillus. More than a hundred species had been identified in the genus paenibacillus, however, few found to cause human infection. This case is unique as it is the first pediatric case with a paenibacillus infection and the first pediatric case where this organism which rarely causes human infection was found in the blood culture of a patient on ECMO.

关键词： paenibacillus ECMO Septic Shock

paenibacillus massiliensis T7固氮基因簇nifBHDKENX的克隆、序列分析及铵和氧对nifB启动子的调控

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中国科学（C辑） 2005年第5期35卷 424-432页

作者：赵洪新刘旭明谢宝恩陈三凤中国农业大学农业生物技术国家重点实验室中国农业大学农业部微生物资源及其应用重点实验室中国农业大学生物学院沈阳师范大学化学与生命科学院沈阳10034

paenibacillus massiliensisT7是我室从北京郊区分离的一株革兰氏阳性、产芽孢的固氮菌.根据GenBank中已发表的多种固氮生物nifH基因序列,在其保守区域设计一对简并引物,PCR扩增获得324bp的nifH片段.用地高辛标记该片段为探针,对EcoRⅠ... 详细信息

paenibacillus massiliensisT7是我室从北京郊区分离的一株革兰氏阳性、产芽孢的固氮菌.根据GenBank中已发表的多种固氮生物nifH基因序列,在其保守区域设计一对简并引物,PCR扩增获得324bp的nifH片段.用地高辛标记该片段为探针,对EcoRⅠ和PstⅠ消化的***7染色体DNA进行Southern杂交,结果DNA/EcoRⅠ杂交道1.1kb和9kb处,DNA/PstⅠ杂交道1.3kb和8kb处分别有两条杂交带,从而初步判断nifH可能有两个拷贝.在324bpnifH片段的基础上,采用反向PCR扩增的方法,获得了purD和nifBHDKENX基因.在nifB的上游有一个类似σ54型启动子的保守序列,在该启动子上游有一个类似激活蛋白NifA结合位点.分析表明,nifBHDKENX可能组成一个转录单元,共用nifB上游的启动子.将nifB启动子与报告基因lacZ相融合,构建成表达载体pnifB-LacZ,并转化到***7中,研究铵和氧对该启动子的表达调控.结果表明,在无氧条件下,铵浓度为0%时,β-半乳糖苷酶的活性最高,随着铵浓度的增加,酶活性并没有明显的下降过程,即使铵浓度达80mmol/L时,酶活也相当于无铵时的73.12%,说明铵对nifB启动子有微弱的抑制作用,而铵浓度一定,氧浓度3%时,β-半乳糖苷酶活性最高,提高氧浓度则酶活性开始下降,当氧浓度高于20%时,β-半乳糖苷酶活性仅几个Miller单位,说明氧对nifB启动子有明显的抑制作用.

关键词： nifBHDKENX 反向PCR 启动子 paenibacillus massiliensis T7 固氮生物表达调控 Southern杂交基因簇

paenibacillus ***3526发酵小麦麸皮生产凝乳酶

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食品与发酵工业 2016年第2期42卷 35-40页

作者：杭锋洪青陶源王钦博刘振民陈卫江南大学食品学院食品科学与技术国家重点实验室江苏无锡214122 乳业生物技术国家重点实验室光明乳业股份有限公司乳业研究院上海200436

对paenibacillus ***3526凝乳酶的发酵条件和凝乳性能进行了研究,探讨了不同培养基、发酵时间、碳源、氮源和装液量对凝乳酶活力和活菌数的影响,并与商业化的小牛皱胃酶、重组凝乳酶、Rhizomucor miehei来源凝乳酶进行凝乳性能比较。实... 详细信息

对paenibacillus ***3526凝乳酶的发酵条件和凝乳性能进行了研究,探讨了不同培养基、发酵时间、碳源、氮源和装液量对凝乳酶活力和活菌数的影响,并与商业化的小牛皱胃酶、重组凝乳酶、Rhizomucor miehei来源凝乳酶进行凝乳性能比较。实验结果表明:发酵时间、碳源、氮源和装液量对BD3526凝乳酶活力(milkclotting activity,MCA)和蛋白水解活力(proteolytic activity,PA)均有不同程度的影响。选用小麦麸皮培养基进行发酵至24 h时,MCA达到最大至3 279.76±67.11 SU/m L,凝乳性能强,PA值在整个发酵期间变化不大;添加碳源或氮源会不同程度地降低MCA值;装液量为30 m L时,发酵上清液中菌体浓度最高、MCA达到6 000 SU/m L。与商业化的凝乳酶相比,BD3256凝乳酶在凝乳过程中未使凝乳块产生过度水解,也无乳清析出现象,具有应用到干酪生产中的潜力。

关键词： paenibacillus spp. 凝乳酶小麦麸皮凝乳酶活力蛋白酶活力