Isolation and characterisation of Methylocystis spp. for poly-3-hydroxybutyrate production using waste methane feedstocks.
Isolation and characterisation of Methylocystis spp. for poly-3-hydroxybutyrate production using waste methane feedstocks.
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DOI：
10.1186/s13568-020-01159-4
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发表时间：
2021-01-06
期刊：
影响因子：
3.7
通讯作者：
Zhang Y
中科院分区：
文献类型：
--
作者：
Rumah BL;Stead CE;Claxton Stevens BH;Minton NP;Grosse-Honebrink A;Zhang Y
废塑料和甲烷排放是加剧环境污染的两种人为副产品。甲烷氧化细菌（甲烷氧化菌）通过利用另外浪费的甲烷气体作为碳源并将碳积累为聚-3-羟基丁酸酯（一种可生物降解的塑料聚合物），同时掌握解决这些问题的关键。在这里，我们提出了两种新的甲基囊藻菌株的分离和表征，当生长在来自不同废物来源（如垃圾填埋场和厌氧消化器气体）的甲烷上时，其能够产生高达细胞干重的55.7 ± 1.9%的聚-3-羟基丁酸酯。利用PacBio和Illumina基因组测序技术对从休闲湖分离的玫瑰甲基囊藻（Methylocystis rosea）BRCS 1和从沼泽分离的小甲基囊藻（Methylocystis parvus）BRCS 2进行全基因组测序。除硝酸钾外，这些菌株还显示在氯化铵、谷氨酰胺和鸟氨酸作为氮源上生长。以0.1%甲醇蒸汽为碳源，在硝酸盐培养基上对小甲基囊藻BRCS 2进行了生长研究。利用带有ColE 1复制起点的质粒，对玫瑰甲基孢囊霉BRCS 1和小甲基孢囊霉BRCS 2的接合效率分别达到2.8 × 10-2和1.8 × 10-2。最后，我们表明，Methylocystis物种可以产生大量的聚-3-羟基丁酸酯的废物甲烷源，而不会损害增长，概念的证明，打开了大门，他们在综合生物设施，如垃圾填埋场和厌氧消化器的使用。
Waste plastic and methane emissions are two anthropogenic by-products exacerbating environmental pollution. Methane-oxidizing bacteria (methanotrophs) hold the key to solving these problems simultaneously by utilising otherwise wasted methane gas as carbon source and accumulating the carbon as poly-3-hydroxybutyrate, a biodegradable plastic polymer. Here we present the isolation and characterisation of two novel Methylocystis strains with the ability to produce up to 55.7 ± 1.9% poly-3-hydroxybutyrate of cell dry weight when grown on methane from different waste sources such as landfill and anaerobic digester gas. Methylocystis rosea BRCS1 isolated from a recreational lake and Methylocystis parvus BRCS2 isolated from a bog were whole genome sequenced using PacBio and Illumina genome sequencing technologies. In addition to potassium nitrate, these strains were also shown to grow on ammonium chloride, glutamine and ornithine as nitrogen source. Growth of Methylocystis parvus BRCS2 on Nitrate Mineral Salt (NMS) media with 0.1% methanol vapor as carbon source was demonstrated. The genetic tractability by conjugation was also determined with conjugation efficiencies up to 2.8 × 10–2 and 1.8 × 10–2 for Methylocystis rosea BRCS1 and Methylocystis parvus BRCS2 respectively using a plasmid with ColE1 origin of replication. Finally, we show that Methylocystis species can produce considerable amounts of poly-3-hydroxybutyrate on waste methane sources without impaired growth, a proof of concept which opens doors to their use in integrated bio-facilities like landfills and anaerobic digesters.
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