Transformation and electrophoretic karyotyping of Coniochaeta ligniaria NRRL30616

Abstract

Coniochaeta ligniaria NRRL30616 is an ascomycete that grows with yeast-like appearance in liquid culture. The strain has potential utility for conversion of fibrous biomass to fuels or chemicals. Furans and other inhibitory compounds in lignocellulosic biomass are metabolized by NRRL30616, facilitating subsequent microbial fermentation of biomass sugars. This study undertook initial characterization of the genetic system of C. ligniaria NRRL30616. Transformation using hygromycin as a dominant selectable marker was achieved using protoplasts generated by incubating cells in 1% (v/v) β-mercaptoethanol, followed by cell wall-digesting enzymes. Thirteen chromosomes with an estimated total size of 30.1 Mb were detected in C. ligniaria. The GC content of chromosomal DNA and of coding regions from cDNA sequences were 49.2 and 51.9%, respectively. This study is the first report of genome size, electrophoretic karyotype, and transformation system for a member of the Coniochaetales.

Production of xylitol by a Coniochaeta ligniaria strain tolerant of inhibitors and defective in growth on xylose

“…Coniochaeta ligniaria C8100 and C8104 are mutants derived from NRRL30616 as follows. Cells of C. ligniaria NRRL30616, an Ascomycete that has yeast‐like growth in liquid culture, were mutagenized by exposure to UV light, then diluted onto solid mineral medium containing 2% (w/v) xylose plus a limiting amount (0.1% w/v) of glucose. This strategy effects a qualitative screen for colonies with normal growth on glucose and impaired growth on xylose.…”

Abatement of Inhibitors in Recycled Process Water from Biomass Fermentations Relieves Inhibition of a Saccharomyces cerevisiae Pentose Phosphate Pathway Mutant

“…Sugars and fermentation products were quantitated using HPLC and refractive index detection [26]. Furfural and HMF in hydrolysate were measured with ultraviolet detection at 277 nm in reverse phase HPLC [13].…”

Effect of lignin-blocking agent on enzyme hydrolysis of acid pretreated hemp waste

“…For example, hot water washing was cost-effective to remove soluble inhibitors, and the addition of activated carbon and a blocking agent (e.g., bovine serum albumin or soybean protein) could reduce enzyme inhibition by lignin or ligninderived compounds. Other studies described various approaches such as liquid-liquid extraction, 46 microbial detoxication, 12,13,47 and genetic modication of low lignin crop 26 to minimize the non-productive binding of enzymes and potential inhibitory effects. Detoxication methods are briey summarized in Table 1B.…”