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Morphology engineering for enhanced production of medium-chain-length polyhydroxyalkanoates in Pseudomonas mendocina NK-01

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Abstract

Polyhydroxyalkanoates (PHAs) can be produced by microorganisms from renewable resources and are regarded as promising bioplastics to replace petroleum-based plastics. A medium-chain-length PHAs (mcl-PHA)-producing strain Pseudomonas mendocina NK-01 was isolated previously by our lab and its whole-genome sequence is currently available. Morphology engineering of manipulating cell morphology–related genes has been applied for enhanced accumulation of the intracellular biopolymer short-chain-length PHAs (scl-PHA). However, it has not yet been reported to improve the yield of mcl-PHA by morphology engineering so far. In this work, several well-characterized cell morphology–related genes, including the cell fission ring (Z-ring) location genes minCD, peptidoglycan degradation gene nlpD, actin-like cytoskeleton protein gene mreB, Z-ring formation gene ftsZ, and FtsZ inhibitor gene sulA, were intensively investigated for their impacts on the cell morphology and mcl-PHA accumulation by gene knockout and overexpression in P. mendocina NKU, a upp knockout mutant of P. mendocina NK-01. For a minCD knockout mutant P. mendocina NKU-∆minCD, the average cell length was obviously increased and the mcl-PHA production was improved. However, the nlpD knockout mutant had a shorter cell length and lower mcl-PHA yield compared with P. mendocina NKU. Overexpression of mreB in P. mendocina NKU resulted in spherical cells. When ftsZ was overexpressed in P. mendocina NKU, the cell division was accelerated and the mcl-PHA titer was improved. Furthermore, mreB, ftsZ, or sulA was overexpressed in P. mendocina NKU-∆minCD. Consequently, the mcl-PHA titers were all increased compared with P. mendocina NKU-∆minCD carrying the empty vector. The multiple fission pattern was finally achieved in ftsZ-overexpressing NKU-∆minCD. In this work, improved production of mcl-PHA in P. mendocina NK-01 has been achieved by morphology engineering. This work provides an alternative strategy to enhance mcl-PHA accumulation in mcl-PHA-producing strains.

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Funding

This work was financially supported by the National Natural Science Funding of China (Grant Nos. 31470213, 31570035 and 31670093), the Tianjin Natural Science Funding (Grant Nos. 17JCZDJC32100 and 18JCYBJC24500), and the Postdoctoral Science Funding of China (Grant No. 2018M631729).

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Authors and Affiliations

  1. Key Laboratory of Molecular Microbiology and Technology for Ministry of Education, Nankai University, Tianjin, 300071, China

    Fengjie Zhao, Ting Gong, Xiangsheng Liu, Xu Fan, Rui Huang, Ting Ma & Chao Yang

  2. State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300071, China

    Shufang Wang & Weixia Gao

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  1. Fengjie Zhao
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  2. Ting Gong
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  3. Xiangsheng Liu
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  4. Xu Fan
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  6. Ting Ma
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  7. Shufang Wang
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  9. Chao Yang
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Correspondence to Shufang Wang, Weixia Gao or Chao Yang.

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Zhao, F., Gong, T., Liu, X. et al. Morphology engineering for enhanced production of medium-chain-length polyhydroxyalkanoates in Pseudomonas mendocina NK-01. Appl Microbiol Biotechnol 103, 1713–1724 (2019). https://doi.org/10.1007/s00253-018-9546-8

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