Screening and breeding of high taxol producing fungi by genome shuffling

Zhao, Kai; Ping, Wenxiang; Zhang, Lína; Liu, Jun; Lin, Yan; Jin, Tao; Zhou, Daohong

doi:10.1007/s11427-008-0037-5

Cited by 37 publications

(24 citation statements)

References 16 publications

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“…74,75 Genome shuffling was applied for breeding the native paclitaxel-producing fungus, Nodulisporum sylviforme strain HQD 33 . 76 First, N. sylviforme spores were treated in several mutagenesis rounds (UV, EMS 60 Co and NTG) to obtain the strain NCEU-1. 77 Next, different combinations of mutagens were applied to the preceding strain to get three hereditarily stable strains named HD 100-23 , UV and UL .…”

Section: Genome Shufflingmentioning

confidence: 99%

Microbial paclitaxel: advances and perspectives

et al. 2010

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Paclitaxel is a potent and widely used antitumor agent. Considerable worldwide research efforts have been carried out on different production alternatives. Since the description of the first paclitaxel-producing fungi, more than 15 years ago, microorganisms have been investigated as potential alternatives for an environmentally acceptable, relatively simple and inexpensive method to produce paclitaxel. However, in spite of significant research on paclitaxel-producing microorganisms, no commercial fermentation process has been implemented up to now. The aim of this study is to review the present status of research on paclitaxel-producing microorganisms and the ongoing efforts to develop heterologous paclitaxel biosynthesis, and analyze the perspectives of microbial fermentation for paclitaxel production.

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Section: Genome Shufflingmentioning

confidence: 99%

Microbial paclitaxel: advances and perspectives

et al. 2010

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“…We therefore used a recursive poolwise mating methodology to shuffle the genomes. A recent study has shown that genome shuffling of S. cerevisiae through recursive mating is possible (15), and this method can circumvent the low efficiency of protoplast fusion (43). The study by Hou increased ethanol tolerance through recombination by means of the S. cerevisiae mating cycle by using a small population of starting mutants for each crossing, a method suitable for evolutionary engineering for tolerance to a single source of inhibition (14).…”

mentioning

confidence: 99%

Saccharomyces cerevisiae Genome Shuffling through Recursive Population Mating Leads to Improved Tolerance to Spent Sulfite Liquor

Pinel

D'Aoust

Cardayré³

et al. 2011

Appl Environ Microbiol

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Spent sulfite liquor (SSL) is a waste effluent from sulfite pulping that contains monomeric sugars which can be fermented to ethanol. However, fermentative yeasts used for the fermentation of the sugars in SSL are adversely affected by the inhibitory substances in this complex feedstock. To overcome this limitation, evolutionary engineering of Saccharomyces cerevisiae was carried out using genome-shuffling technology based on large-scale population cross mating. Populations of UV-light-induced yeast mutants more tolerant than the wild type to hardwood spent sulfite liquor (HWSSL) were first isolated and then recursively mated and enriched for more-tolerant populations. After five rounds of genome shuffling, three strains were isolated that were able to grow on undiluted HWSSL and to support efficient ethanol production from the sugars therein for prolonged fermentation of HWSSL. Analyses showed that greater HWSSL tolerance is associated with improved viability in the presence of salt, sorbitol, peroxide, and acetic acid. Our results showed that evolutionary engineering through genome shuffling will yield robust yeasts capable of fermenting the sugars present in HWSSL, which is a complex substrate containing multiple sources of inhibitors. These strains may not be obtainable through classical evolutionary engineering and can serve as a model for further understanding of the mechanism behind simultaneous tolerance to multiple inhibitors.

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“…PuriWed YX-3 protoplast suspensions were transferred into sterile test tubes and incubated in a 60°C water bath for 2, 7, 12, 17, 22, and 27 min to choose the optimal inactivation condition [28,30]. Inactivation was conWrmed by lack of growth in the YPDS medium.…”

Section: Protoplast Inactivationmentioning

confidence: 99%

“…The puriWed LX-4 protoplast suspensions were transferred to a sterile Petri dish (6 cm diameter) which was then placed under a preheated 30-W UV lamp at a vertical distance of 20 cm and irradiated for 10, 20, 30, 40, 45, and 50 min to choose the optimal inactivation condition [3,24,28]. The treated protoplasts were maintained in the dark for 2 h to avoid photoreactivation repair.…”

Section: Protoplast Inactivationmentioning

confidence: 99%

“…Activation in the presence of LiCl was carried out by adding LiCl solution to a puriWed CXS-5 protoplast suspension to a Wnal concentration of 0.6% (w/v) and then irradiated for 5, 10, 15, 20, 25, and 30 min under a preheated 30-W UV lamp at a vertical distance of 20 cm to choose the optimal inactivation condition [28]. The treated protoplasts were kept in the dark for 2 h to avoid photoreactivation repair.…”

Section: Protoplast Inactivationmentioning

confidence: 99%

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A genome shuffling-generated Saccharomyces cerevisiae isolate that ferments xylose and glucose to produce high levels of ethanol

Sun

Song

et al. 2012

Journal of Industrial Microbiology and Biotechnology

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Genome shuffling is an efficient approach for the rapid improvement of industrially important microbial phenotypes. This report describes optimized conditions for protoplast preparation, regeneration, inactivation, and fusion using the Saccharomyces cerevisiae W5 strain. Ethanol production was confirmed by TTC (triphenyl tetrazolium chloride) screening and high-performance liquid chromatography (HPLC). A genetically stable, high ethanol-producing strain that fermented xylose and glucose was obtained following three rounds of genome shuffling. After fermentation for 84 h, the high ethanol-producing S. cerevisiae GS3-10 strain (which utilized 69.48 and 100% of the xylose and glucose stores, respectively) produced 26.65 g/L ethanol, i.e., 47.08% higher than ethanol production by S. cerevisiae W5 (18.12 g/L). The utilization ratios of xylose and glucose were 69.48 and 100%, compared to 14.83 and 100% for W5, respectively. The ethanol yield was 0.40 g/g (ethanol/consumed glucose and xylose), i.e., 17.65% higher than the yield by S. cerevisiae W5 (0.34 g/g).

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Screening and breeding of high taxol producing fungi by genome shuffling

Cited by 37 publications

References 16 publications

Microbial paclitaxel: advances and perspectives

Microbial paclitaxel: advances and perspectives

Saccharomyces cerevisiae Genome Shuffling through Recursive Population Mating Leads to Improved Tolerance to Spent Sulfite Liquor

A genome shuffling-generated Saccharomyces cerevisiae isolate that ferments xylose and glucose to produce high levels of ethanol

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