Biodegradation of High Molecular Weight Alkanes by the Microbiome of Galleria mellonella Larvae

大蜡螟幼虫微生物组对高分子量烷烃的生物降解

基本信息

  • 批准号:
    RGPIN-2018-06166
  • 负责人:
  • 金额:
    $ 1.89万
  • 依托单位:
  • 依托单位国家:
    加拿大
  • 项目类别:
    Discovery Grants Program - Individual
  • 财政年份:
    2020
  • 资助国家:
    加拿大
  • 起止时间:
    2020-01-01 至 2021-12-31
  • 项目状态:
    已结题

项目摘要

This proposal is designed to train at least 2 PhD, 3 MSc and 15 UG students in a multidisciplinary program. Alkanes longer than nonane are considered to be long chain alkanes. While microorganisms have been isolated that can degrade C20-C50 alkanes, the degradation rate is very low and the mechanism of degradation is unclear. This proposal focuses on the study of the degradation of C20 or longer linear and branched alkanes by greater waxmoth (Galleria mellonella) larvae and microorganisms from its digestive tract. The research is based on the hypothesis that polyethylene (PE)-biodegraders can also degrade long chain alkanes. PEs are high molecular weight alkanes of varying branching and crystallinity. Recently, it was demonstrated that larvae of G. mellonella and related organisms can degrade PE and PE-degrading bacteria have been isolated from their gut. G. mellonella larvae will be grown in the lab using PE and, for the first time, a variety of linear and branched alkanes both to study the range of alkanes that can be degraded by these larvae and to generate inocula (ground-up digestive tracts and fecula) for microbial enrichment cultures. Under anaerobic conditions there is preferential loss of high molecular weight n-alkanes in petroleum-contaminated sites, and there is preferential degradation of branched over normal alkanes under sulphate reducing conditions. Therefore, enrichment culture conditions employed will be aerobic and, for the first time using G. mellonella inocula, anaerobic conditions with a variety of terminal electron acceptors. A range of 13C alkane substrates will be synthesized and spiked into microcosms under aerobic, nitrate-, sulphate-reducing and methanotrophic conditions to identify alkane degraders. Total community DNA will be recovered and separated by density gradient centrifugation to recover the degrading population with 13C-enriched DNA. PCR amplification and purification, and Illumina MiSeq sequencing and data processing will be employed to identify the degraders. The abilities of isolates to degrade a range of alkanes of different molecular weights and branching will be examined in pure culture. Degradation products, DSC analysis of crystallinity changes and surface tension data of liquid media will provide insight into degradation mechanisms. The proposed program sets the framework for a series of investigations into biodegradation of plastics and petrochemicals based on the microbiome of G. mellonella and related organisms. This work will add significantly to our knowledge of the possibilities for biodegradation of recalcitrant petroleum fractions and plastics. Identification and isolation of degraders creates the opportunity to use these organisms in bioaugmentation, as well as informing engineers as to the appropriate physiological conditions (redox, electron acceptor) required for degradation of recalcitrant alkanes.
该提案旨在培养至少2名博士,3名硕士和15名UG学生的多学科课程。比壬烷长的烷烃被认为是长链烷烃。虽然已经分离出能够降解C20-C50烷烃的微生物,但是降解率非常低,并且降解机理不清楚。该提案的重点是研究大蜡螟(Galleria mellonella)幼虫及其消化道微生物对C20或更长的直链和支链烷烃的降解。该研究基于聚乙烯(PE)生物降解剂也可以降解长链烷烃的假设。PE是具有不同支化度和结晶度的高分子量烷烃。最近的研究表明,在中国大陆,G.大蜡螟及其相关微生物可以降解PE,并且已经从它们的肠道中分离出PE降解细菌。G.将在实验室中使用PE和首次使用各种直链和支链烷烃培养大蜡螟幼虫,以研究可以被这些幼虫降解的烷烃的范围,并产生用于微生物富集培养的接种物(磨碎的消化道和粪)。在厌氧条件下,有优先损失的高分子量的正构烷烃在石油污染的网站,并有优先降解的支链烷烃在硫酸盐还原条件下。因此,所用的富集培养条件将是需氧的,并且首次使用G.大蜡螟接种物,厌氧条件下的各种终端电子受体。将合成一系列13 C烷烃底物,并在需氧、硝酸盐、硫酸盐还原和甲烷氧化条件下掺入微宇宙中,以确定烷烃降解剂。将回收总群落DNA,并通过密度梯度离心分离,以回收具有13 C富集DNA的降解群体。将采用PCR扩增和纯化以及Illumina MiSeq测序和数据处理来鉴定降解物。 将在纯培养物中检查分离物降解一系列不同分子量和分支的烷烃的能力。降解产物、结晶度变化的DSC分析和液体介质的表面张力数据将提供对降解机制的深入了解。该计划为基于G.梅隆菌和相关生物。这项工作将大大增加我们的知识的可能性生物降解的难降解石油馏分和塑料。降解剂的鉴定和分离为在生物强化中使用这些生物体创造了机会,并告知工程师降解柠檬酸烷烃所需的适当生理条件(氧化还原,电子受体)。

项目成果

期刊论文数量(0)
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Ramsay, Bruce其他文献

Carbon-limited fed-batch production of medium-chain-length polyhydroxyalkanoates by a phaZ-knockout strain of Pseudomonas putida KT2440
Increasing the yield of MCL-PHA from nonanoic acid by co-feeding glucose during the PHA accumulation stage in two-stage fed-batch fermentations of Pseudomonas putida KT2440
  • DOI:
    10.1016/j.jbiotec.2007.02.023
  • 发表时间:
    2007-11-01
  • 期刊:
  • 影响因子:
    4.1
  • 作者:
    Sun, Zhiyong;Ramsay, Juliana A.;Ramsay, Bruce
  • 通讯作者:
    Ramsay, Bruce
Enhanced yield of medium-chain-length polyhydroxyalkanoates from nonanoic acid by co-feeding glucose in carbon-limited, fed-batch culture
  • DOI:
    10.1016/j.jbiotec.2009.07.014
  • 发表时间:
    2009-09-25
  • 期刊:
  • 影响因子:
    4.1
  • 作者:
    Sun, Zhiyong;Ramsay, Juliana;Ramsay, Bruce
  • 通讯作者:
    Ramsay, Bruce
Decaying exponential feeding of nonanoic acid for the production of medium-chain-length poly(3-hydroxyalkanoates) by Pseudomonas putida KT2440

Ramsay, Bruce的其他文献

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{{ truncateString('Ramsay, Bruce', 18)}}的其他基金

Biodegradation of High Molecular Weight Alkanes by the Microbiome of Galleria mellonella Larvae
大蜡螟幼虫微生物组对高分子量烷烃的生物降解
  • 批准号:
    RGPIN-2018-06166
  • 财政年份:
    2022
  • 资助金额:
    $ 1.89万
  • 项目类别:
    Discovery Grants Program - Individual
Biodegradation of High Molecular Weight Alkanes by the Microbiome of Galleria mellonella Larvae
大蜡螟幼虫微生物组对高分子量烷烃的生物降解
  • 批准号:
    RGPIN-2018-06166
  • 财政年份:
    2021
  • 资助金额:
    $ 1.89万
  • 项目类别:
    Discovery Grants Program - Individual
Biodegradation of High Molecular Weight Alkanes by the Microbiome of Galleria mellonella Larvae
大蜡螟幼虫微生物组对高分子量烷烃的生物降解
  • 批准号:
    RGPIN-2018-06166
  • 财政年份:
    2019
  • 资助金额:
    $ 1.89万
  • 项目类别:
    Discovery Grants Program - Individual
Biodegradation of High Molecular Weight Alkanes by the Microbiome of Galleria mellonella Larvae
大蜡螟幼虫微生物组对高分子量烷烃的生物降解
  • 批准号:
    RGPIN-2018-06166
  • 财政年份:
    2018
  • 资助金额:
    $ 1.89万
  • 项目类别:
    Discovery Grants Program - Individual
Control of medium-chain-length PHA properties
控制中链长度 PHA 特性
  • 批准号:
    25469-2009
  • 财政年份:
    2013
  • 资助金额:
    $ 1.89万
  • 项目类别:
    Discovery Grants Program - Individual
Control of medium-chain-length PHA properties
控制中链长度 PHA 特性
  • 批准号:
    25469-2009
  • 财政年份:
    2012
  • 资助金额:
    $ 1.89万
  • 项目类别:
    Discovery Grants Program - Individual
Control of medium-chain-length PHA properties
控制中链长度 PHA 特性
  • 批准号:
    25469-2009
  • 财政年份:
    2011
  • 资助金额:
    $ 1.89万
  • 项目类别:
    Discovery Grants Program - Individual
Control of medium-chain-length PHA properties
控制中链长度 PHA 特性
  • 批准号:
    25469-2009
  • 财政年份:
    2010
  • 资助金额:
    $ 1.89万
  • 项目类别:
    Discovery Grants Program - Individual
Control of medium-chain-length PHA properties
控制中链长度 PHA 特性
  • 批准号:
    25469-2009
  • 财政年份:
    2009
  • 资助金额:
    $ 1.89万
  • 项目类别:
    Discovery Grants Program - Individual
Development of mcl-PHA as a bioactive scaffold for tissue engineering
mcl-PHA作为组织工程生物活性支架的开发
  • 批准号:
    25469-2006
  • 财政年份:
    2008
  • 资助金额:
    $ 1.89万
  • 项目类别:
    Discovery Grants Program - Individual

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Biodegradation of High Molecular Weight Alkanes by the Microbiome of Galleria mellonella Larvae
大蜡螟幼虫微生物组对高分子量烷烃的生物降解
  • 批准号:
    RGPIN-2018-06166
  • 财政年份:
    2022
  • 资助金额:
    $ 1.89万
  • 项目类别:
    Discovery Grants Program - Individual
Biodegradation of High Molecular Weight Alkanes by the Microbiome of Galleria mellonella Larvae
大蜡螟幼虫微生物组对高分子量烷烃的生物降解
  • 批准号:
    RGPIN-2018-06166
  • 财政年份:
    2021
  • 资助金额:
    $ 1.89万
  • 项目类别:
    Discovery Grants Program - Individual
Biodegradation of High Molecular Weight Alkanes by the Microbiome of Galleria mellonella Larvae
大蜡螟幼虫微生物组对高分子量烷烃的生物降解
  • 批准号:
    RGPIN-2018-06166
  • 财政年份:
    2019
  • 资助金额:
    $ 1.89万
  • 项目类别:
    Discovery Grants Program - Individual
Biodegradation of High Molecular Weight Alkanes by the Microbiome of Galleria mellonella Larvae
大蜡螟幼虫微生物组对高分子量烷烃的生物降解
  • 批准号:
    RGPIN-2018-06166
  • 财政年份:
    2018
  • 资助金额:
    $ 1.89万
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污染土壤中高分子量多环芳烃的生物降解
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Biodegradation of high molecular weight PAHs in contaminated soil
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