Protoplast Fusion in Streptomyces sp. for Increased Production of Laccase and Associated Ligninolytic Enzymes

Zhan

Env Prog and Sustain Energy

et al. 2012

Protoplast fusion is an important technique for engineering microbial strains. It has been used to combine genes from different organisms to create new strains with desired properties. This investigation was aimed to construct a new strain with an ability of degrading chlorophenols efficiently by protoplast fusion between Pseudomonas putida and Psathyrella candolleana. Twenty recombinant strains were screened and used to degrade pentachlorophenol (PCP) synthetic wastewater in order to determine their capabilities of chlorophenol degradation. To rapidly and accurately measure PCP content in synthetic wastewater after treatment by different strains, a new method quantifying PCP was proposed based on derivative spectroscopy. Chlorophenol biodegradability of different strains could be measured rapidly according to the new method. The results showed that PCP degradation % of four recombinant strains viz., Xz 6‐1, Xz 6‐3, Xz 6‐5, Xz 8‐2 were 78.98%, 69.12%, 31.40%, 46.26%, respectively, and that of parent strain (Pseudomonas putida.) was 57.27%, which indicated that the degradation % of Xz 6‐1 is the highest and improved by 21.71% compared with Pseudomonas putida. Furthermore, the colony morphologies and microscopic feature of Xz 6‐1 and Pseudomonas putida were compared in this article. © 2012 American Institute of Chemical Engineers Environ Prog, 32: 443–448, 2013

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Construction of engineering microorganism degrading chlorophenol efficiently by protoplast fusion technique

Zhan

Env Prog and Sustain Energy

et al. 2012

World J Microbiol Biotechnol

“…A functional strain was also constructed through intraspecific protoplast fusion of Candida tropicalis to enhance phenol degradation (Chang et al 1995). Sivakumar et al (2004) constructed a protoplast fusant in Streptomyces sp. to degradate lignin.…”

Section: Discussionmentioning

confidence: 99%

Identification of protoplast fusion strain Fhhh by randomly amplified polymorphic DNA

Zhao

Zhang

et al. 2009

A functional strain Fhhh was constructed through protoplast fusion of three parental strains (Phanerochaete chrysosporium, Saccharomyces cerevisiae and native bacterium YZ) to improve the degradation efficiency of purified terephthalic acid wastewater. Randomly amplified polymorphic DNA (RAPD) and scanning electron microscope (SEM) analysis were applied to identify and confirm the fusant Fhhh through phenotypic and genetic relationship. The result of SEM analysis demonstrated that the cell shape of fusant Fhhh differed from all three parental strains. RAPD analysis of 40 arbitrary primers generated a total of 1,135 bands. The genetic similarity indices between Fhhh and parental strains Phanerochaete chrysosporium (PC), Saccharomyces cerevisiae (SC) and native bacterium (YZ) were 34.01%, 33.16%, and 35.97%, respectively. The targeted-gene PCR results showed that Fhhh inherited the DNA fragments of mnp and lip genes from parental strain PC and FLO1 gene fragment from parental strain SC. Our results suggested protoplast fusion technique may be considered as a promising technique in environmental pollution control.

“…Appropriate concentration of osmoticum can prevent the protoplast from bursting or shrinking. Though there is no uniform standard for the selection of a suitable osmotic stabilizer [10,15], it was demonstrated in this study that mannitol was a good choice for P. ostreatus. The best results were achieved with the 0.6M mannitol osmotic stabilizer with protoplast formation and regeneration of P. ostreatus of 4.13×10 8 and 5.67%, respectively …”

Section: The Effect Of Osmotic Stabilizer On Protoplast Formation Andmentioning

confidence: 99%

High-Yield Laccase-Producing Strains Constructed by Protoplast Fusion Between Bacterium and Fungus

Zhao¹,

Wu²,

Wang³

2015

TOBIOTJ

This study is based on the construction of high-yield laccase-producing fusant achieved by inter-kingdom protoplast fusion between Pleurotus ostreatus and Escherichia coli. The optimized protoplasts formation and regeneration conditions were demonstrated with the presence of 1.5% cellulase +1.0% snailase and 0.6M mannitol at 30 o C for 3h. The fusants were screened for different characteristics between two parental strains and further identified by laccase activity, offering one of the genetically stable fusants, Strain F. The fusant F produced the highest yield of laccase, being about 22% higher than that of the parental strain. The results suggest that the protoplast fusion technique can be considered as a promising technique for the control of environmental pollution.