Name: production of chemicals by klebsiella pneumoniae using bamboo hydrolysate as feedstock
Uploaded: 2017-06-29T14:00:00
Description: Watch this Scientific Journal Video about Production of Chemicals by Klebsiella pneumoniae Using Bamboo Hydrolysate as Feedstock at JoVE.com

这项工作得到了国家自然科学基金（批准号：21576279,20906076）和KRIBB研究计划（KGM2211531）的支持。

竹水合物的制备<ol><li> 在40 mL NaOH溶液中加入5 g竹粉，使其在250 mL烧瓶中达到10％（g / g）终浓度。使用0.05M至0.50M的一系列NaOH溶液，以0.05M的增量。 <ol><li>将混合物在60℃或100℃的水浴中孵育60分钟。在121℃下在高压釜中孵育。 </li></ol></li><li> 孵育后，将混合物在室温下静置4小时。然后，取出上清液。加入100ml淡水并混合。 <ol><li>重复洗涤5-6次，直到上清液的pH达到6.8。使?...

<ol>
	<li>Zeng, A. P., Biebl, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bulk+chemicals+from+biotechnology:+the+case+of+1,+3-propanediol+production+and+the+new+trends.">Bulk chemicals from biotechnology: the case of 1, 3-propanediol production and the new trends.</a> Adv Biochem Eng Biotechnol. 74, 239-259 (2002).</li><li>Wei, D., Wang, M., Jiang, B., Shi, J., Hao, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+dihydroxyacetone+kinases+I+and+II+in+the+dha+regulon+of+Klebsiella+pneumoniae.">Role of dihydroxyacetone kinases I and II in the dha regulon of Klebsiella pneumoniae.</a> J Biotechnol. 177, 13-19 (2014).</li><li>Celi&#324;ska, E., Grajek, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biotechnological+production+of+2,3-butanediol-current+state+and+prospects.">Biotechnological production of 2,3-butanediol-current state and prospects.</a> Biotechnol Adv. 27 (6), 715-725 (2009).</li><li>Chen, C., Wei, D., Shi, J., Wang, M., Hao, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanism+of+2,+3-butanediol+stereoisomer+formation+in+Klebsiella+pneumoniae.">Mechanism of 2, 3-butanediol stereoisomer formation in Klebsiella pneumoniae.</a> Appl Microbiol Biotechnol. 98 (10), 4603-4613 (2014).</li><li>Wei, D., Wang, M., Shi, J., Hao, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Red+recombinase+assisted+gene+replacement+in+Klebsiella+pneumoniae.">Red recombinase assisted gene replacement in Klebsiella pneumoniae.</a> J Ind Microbiol Biotechnol. 39 (8), 1219-1226 (2012).</li><li>Wei, D., Sun, J., Shi, J., Liu, P., Hao, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+strategy+to+improve+efficiency+for+gene+replacement+in+Klebsiella+pneumoniae.">New strategy to improve efficiency for gene replacement in Klebsiella pneumoniae.</a> J Ind Microbiol Biotechnol. 40 (5), 523-527 (2013).</li><li>Chen, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+RecBCD+in+Klebsiella+pneumoniae+by+Gam+and+its+effect+on+the+efficiency+of+gene+replacement.">Inhibition of RecBCD in Klebsiella pneumoniae by Gam and its effect on the efficiency of gene replacement.</a> J Basic Microbiol. 56 (2), 120-126 (2016).</li><li>Wang, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=R-acetoin+accumulation+and+dissimilation+in+Klebsiella+pneumoniae.">R-acetoin accumulation and dissimilation in Klebsiella pneumoniae.</a> J Ind Microbiol Biotechnol. 42 (8), 1105-1115 (2015).</li><li>Wei, D., Xu, J., Sun, J., Shi, J., Hao, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=2-Ketogluconic+acid+production+by+Klebsiella+pneumoniae+CGMCC+1.6366.">2-Ketogluconic acid production by Klebsiella pneumoniae CGMCC 1.6366.</a> J Ind Microbiol Biotechnol. 40 (6), 561-570 (2013).</li><li>Sun, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two-stage+fermentation+for+2-ketogluconic+acid+production+by+Klebsiella+pneumoniae.">Two-stage fermentation for 2-ketogluconic acid production by Klebsiella pneumoniae.</a> J Microbiol Biotechnol. 24 (6), 781-787 (2014).</li><li>Wang, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gluconic+acid+production+by+gad+mutant+of+Klebsiella+pneumoniae.">Gluconic acid production by gad mutant of Klebsiella pneumoniae.</a> World J Microbiol Biotechnol. 32 (8), 1-11 (2016).</li><li>Wang, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Production+of+xylonic+acid+by+Klebsiella+pneumoniae.">Production of xylonic acid by Klebsiella pneumoniae.</a> Appl Microbiol Biotechnol. 100 (23), 10055-10063 (2016).</li><li>Kumar, P., Barrett, D. M., Delwiche, M. J., Stroeve, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+for+pretreatment+of+lignocellulosic+biomass+for+efficient+hydrolysis+and+biofuel+production.">Methods for pretreatment of lignocellulosic biomass for efficient hydrolysis and biofuel production.</a> Ind Eng Chem Res. 48 (8), 3713-3729 (2009).</li><li>Brisse, S., Grimont, F., Grimont, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genus+Klebsiella.">The genus Klebsiella.</a> The Prokaryotes. , 159-196 (2006).</li><li>Hao, J., Lin, R., Zheng, Z., Liu, H., Liu, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+characterization+of+microorganisms+able+to+produce+1,+3-propanediol+under+aerobic+conditions.">Isolation and characterization of microorganisms able to produce 1, 3-propanediol under aerobic conditions.</a> World J Microbiol Biotechnol. 24 (9), 1731-1740 (2008).</li><li>Hong, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimization+of+alkaline+pretreatment+on+corn+stover+for+enhanced+production+of+1.3-propanediol+and+2,+3-butanediol+by+Klebsiella+pneumoniae+AJ4.">Optimization of alkaline pretreatment on corn stover for enhanced production of 1.3-propanediol and 2, 3-butanediol by Klebsiella pneumoniae AJ4.</a> Biomass Bioenerg. 77, 177-185 (2015).</li><li>Pienkos, P. T., Zhang, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+pretreatment+and+conditioning+processes+on+toxicity+of+lignocellulosic+biomass+hydrolysates.">Role of pretreatment and conditioning processes on toxicity of lignocellulosic biomass hydrolysates.</a> Cellulose. 16 (4), 743-762 (2009).</li></ol>

肺炎 克雷伯杆菌属于肠杆菌科科克雷伯 菌属 。 肺炎克雷伯病在土壤，植被和水等自然环境中广泛分布14 。本研究中使用的野生型肺炎克雷伯杆菌菌株从土壤中分离，用​​于1,3-丙二醇生产15 。这种物种的肺炎支原体和突变体在不同条件下产生许多化学物质。 培养pH和空气补充是影响肺?...

肺炎克雷伯杆菌是一种在有氧和无氧条件下都能良好生长的细菌。 肺炎支原体是用于生产许多化学物质的重要工业微生物。 1,3-丙二醇是一种有价值的化学物质，主要用作合成聚对苯二甲酸丙二醇酯的单体。聚对苯二甲酸丙二醇酯是一种可生物降解的聚酯，其表现出比1,2-丙二醇，丁二醇或乙二醇1更好的性能。 1,3-丙二醇由肺炎链球菌在氧气限制条件下使用甘油作为底物生产2 。 2,3-丁二醇及其衍生物在塑料，溶剂生产和合成橡胶领域有应用，有潜力被用作生物燃料3 。以葡萄糖为底物，2,3-丁二醇是野生型菌株4的主要代谢物。 2,3-丁二醇是合成的从丙酮酸。首先，两分子丙酮酸缩合得到α-乙酰乳酸;该反应由α-乙酰乳酸合成酶催化。然后通过α-乙酰乳酸脱羧酶将α-乙酰乳酸转变成乙偶姻。当丁二醇脱氢酶催化时，β-维生素可进一步还原成2,3-丁二醇。已经探索了适用于肺炎克雷伯菌的有效基因替代方法，并且已经构建了许多突变体5,6,7 。失去其2,3-丁二醇脱氢酶活性的budC突变体在培养液中积累高水平的乙偶姻。乙酰头痛被用作增加食物味道的添加剂。当编码α-乙酰乳酸脱羧酶的budA被突变时，2-ketogluconic acid积聚在肉汤中。 2-酮基葡萄糖酸用于合成异抗坏血酸（异抗坏血酸），用于食品工业的抗氧化剂9 。 2-酮葡萄糖酸是葡萄糖氧化途径的中间体;在该通路中，位于周质空间中，葡萄糖被氧化成葡萄糖酸，然后进一步氧化成2-酮基葡萄糖酸。葡萄糖酸和在周质中产生的2-酮葡萄糖酸可以转运到细胞质进一步代谢。 2-酮基葡萄糖酸的积累取决于酸性条件，更高的空气补充有利于2-酮葡萄糖酸生产10 。葡萄糖酸脱氢酶，由gad编码， 催化葡萄糖酸转化为2-酮聚糖酸。 肺炎克雷伯氏菌的gad突变体产生高水平的葡萄糖酸而不是2-酮葡萄糖酸，并且该过程也依赖于酸性条件。葡萄糖酸是一种大量有机酸，用作添加剂以增加水泥1的性能1。葡萄糖氧化成葡萄糖酸由葡萄糖脱氢酶催化。木糖也是葡萄糖脱氢酶的合适底物。当木糖用作底物时， 肺炎克雷伯菌产生木糖酸12 。 使用生物质作为原料的化学生产是生物技术的热门话题13 。生物质的主要成分是纤维素，半纤维素和木质素。然而，这些大分子化合物不能被大多数微生物（包括肺炎克雷伯菌 ）直接分解代谢。生物质中的纤维素和半纤维素必须水解成葡萄糖和木糖，然后可以被微生物使用。在木质纤维素中木质素的存在产生防止酶的生物质水解的保护屏障。因此，消除木质素和半纤维素并降低纤维素的结晶度的预处理过程总是在通过麦克风的生物量利用期间进行roorganisms。已经开发了许多预处理方法：酸，碱，氨和蒸汽预处理是常见的。 竹子在热带和亚热带地区丰富，是重要的生物质资源。本文介绍了竹水解产物的制备和竹水解产物的化学生产

<table border="0" cellpadding="0" cellspacing="0" width="638">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">autoclave</td>
			<td style="width:136px;">SANYO</td>
			<td align="right" style="width:119px;">3780</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">bioreactor</td>
			<td style="width:136px;">Sartorius stedim biotech</td>
			<td style="width:119px;">Bostat Aplus</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">agitator</td>
			<td style="width:136px;">IKA</td>
			<td style="width:119px;">RW 20</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">water bath shaker</td>
			<td style="width:136px;">Zhicheng</td>
			<td style="width:119px;">ZWY-110X50</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">high performance liquid chromatograph system</td>
			<td style="width:136px;">Shimadzu Corp</td>
			<td style="width:119px;">20AVP</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">centrifuge</td>
			<td style="width:136px;">Hitachi</td>
			<td style="width:119px;">CR22G III</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Bamboo powder</td>
			<td style="width:136px;">purchased from Zhejiang Province, China</td>
			<td style="width:119px;"></td>
			<td>mesh number, 50</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">cellulase</td>
			<td style="width:136px;">Youtell Biochemical, Shandong, China</td>
			<td style="width:119px;"></td>
			<td>200 PFU/ml</td>
		</tr>
	</tbody>
</table>

production of chemicals by klebsiella pneumoniae using bamboo hydrolysate as feedstock

竹是重要的生物质，竹科水解产物被克雷伯杆菌用作化学生产的原料。这里，用NaOH预处理竹粉，洗涤至中性pH。将纤维素酶加入预处理的竹粉中以产生含有30g / L葡萄糖和15g / L木糖的水解产物，并用作碳源以制备用于化学生产的培养基。当在微需氧条件下培养时，野生型肺炎克雷伯菌产生12.7g / L 2,3-丁二醇。在需氧条件下， 肺炎支原体 芽胞芽孢杆菌突变体产生了13.0g / L的R-丝氨酸。在肺炎链球菌的budA突变体的两阶段，pH控制的发酵中补充高空气产生了25.5g / L的2-酮基葡萄糖酸和13.6g / L的木糖酸的混合物。在发酵的第一阶段，将培养物保持在中性pH值;细胞生长后，发酵n进入第二阶段，在此阶段，文化被允许变成酸性。

在这个方案中，竹子是用碱预处理的。在图1和图 2中确定了最佳孵育参数 - 温度为121℃和0.25M NaOH。将预处理的竹子酶水解，测定水解产物中得到的葡萄糖和木糖浓度。更高的温度有利于糖的生产，所以选择121°C作为最佳温度。在水解产物中产生的葡萄糖和木糖在NaOH浓度范围内在0.05-0.25Ma范围内增加。NaOH浓度的进一步增加对糖的产生?...

Watch this Scientific Journal Video about Production of Chemicals by Klebsiella pneumoniae Using Bamboo Hydrolysate as Feedstock at JoVE.com

Production of Chemicals by Klebsiella pneumoniae Using Bamboo Hydrolysate as Feedstock

竹粉用NaOH预处理，酶水解。竹纤维素水解产物被用作2,3-丁二醇，β-乙酰胆碱，2-酮基葡萄糖酸和肺炎克雷伯菌产生的木糖酸的原料。

化学品生产<em&gt;肺炎克雷伯菌</em使用竹水合物作为原料

Bamboo is an important biomass, and bamboo hydrolysate is used by Klebsiella pneumoniae as a feedstock for chemical production. Here, bamboo powder was ...

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Bioengineering

Production of Chemicals by Klebsiella pneumoniae Using Bamboo Hydrolysate as Feedstock

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When present in pharmaceutical products, a Gram-negative bacterial cell wall component endotoxin (often also called lipopolysaccharide) can cause ...

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The presented protocol enables a high-throughput continuous preparation of low temperature-sensitive liposomes (LTSLs), which are capable of loading ...