Enhanced phenolic compounds tolerance response of Clostridium beijerinckii NCIMB 8052 by inactivation of Cbei_3304

Abstract

BackgroundPhenolic compounds generated in hydrolysis of lignocellulosic materials are major limiting factors for biological production of solvents by Clostridia, but it lacks the attention on the study of adaptation or resistance mechanisms in response to phenolic compounds.ResultsGene Cbei_3304, encoding a hypothetical membrane transport protein, was analyzed by bioinformatic method. After insertional inactivation of the functionally uncertain gene Cbei_3304 in Clostridium beijerinckii NCIMB 8052, resulted in enhanced phenolic compounds tolerance. Compared to the parent strain C. beijerinckii NCIMB 8052, evaluation of toxicity showed the recombination stain C. beijerinckii 3304::int had a higher level of tolerance to four model phenolic compounds of lignocellulose-derived microbial inhibitory compounds. A comparative transcriptome analysis showed that the genes were involved in membrane transport proteins (ABC and MFS family) and were up-regulated expression after disrupting gene Cbei_3304. Additionally, the adaptation of C. beijerinckii NCIMB 8052 in response to non-detoxified hemicellulosic hydrolysate was improved by disrupting gene Cbei_3304.ConclusionToxicity evaluation of lignocellulose-derived phenolic compounds shows that Cbei_3304 plays a significant role in regulating toxicities tolerance for ABE fermentation by C. beijerinckii, and the adaptation of non-detoxified hemicellulosic hydrolysate is significantly improved after inactivation of Cbei_3304 in wild-type strain C. beijerinckii NCIMB 8052. It provided a potential strategy for generating high inhibitor tolerance strains for using lignocellulosic materials to produce solvents by clostridia in this study.Electronic supplementary materialThe online version of this article (10.1186/s12934-018-0884-0) contains supplementary material, which is available to authorized users.

Combined evolutionary engineering and genetic manipulation improve low pH tolerance and butanol production in a synthetic microbial Clostridium community

“…The reason is that the descendants of 743BΔspo0A exhibited better evolutionary fitness than those of the wild type (Wen et al, 2019). Moreover, this was expected to obtain a mutant to tolerate low pH, using novel stress‐resistance molecular machines (such as two‐component systems or membrane transport protein; J. Liu, Lin, Chai, Luo, & Guo, 2018; Xu et al, 2015), but not by known mechanisms, such as spore formation or the regulation of spo0A expression levels (thus regulating target genes; Al‐Hinai, Jones, & Papoutsakis, 2015).…”

Improved n -Butanol Production from Clostridium cellulovorans by Integrated Metabolic and Evolutionary Engineering

“…We hypothesized that there may be further molecular mechanisms that lead to stress resistance in C. cellulovorans, such as several two-component systems or membrane transport proteins, that are not entirely dependent on spo0A (53,54). When spo0A is knocked out, attenuated, or mutated, other mechanisms may be activated or reinforced for the strain to respond to butanol stress.…”

TargeTron Technology Applicable in Solventogenic Clostridia: Revisiting 12 Years’ Advances

“…Above method also was applied to the functional analysis of other important regulatory genes, such as Rex (CAC2713) [77,81], celR (CAC0382) [82], AbrB (AbrB0310 and AbrB3647) [83], csrA (CAC2209) [84], etc. Besides, TargeTron plays a key role in identification of some other important functional genes such as the stress resistance genes CAC3319 and Cbei3304 [85,86] or the quorum sensing gene agr ABC (CAC0078-0081) [87]. Generally, TargeTron has become an important genetic tool for gene function identification.…”