Pedrino Matheus, Narcizo Julia Pereira, Aguiar Inaiá Ramos, Reginatto Valeria, Guazzaroni María-Eugenia
Department of Biology, Faculty of Philosophy, Sciences and Letters of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo 14040-901, Brazil.
Department of Chemistry, Faculty of Philosophy, Sciences and Letters of Ribeirão Preto, University of São Paulo, Ribeirão Preto 14040-900, Brazil.
ACS Omega. 2025 Feb 10;10(6):5449-5459. doi: 10.1021/acsomega.4c07288. eCollection 2025 Feb 18.
Several species, including KT2440, have a broad metabolic repertoire to assimilate biomass monomers such as lignin-derived compounds but struggle to tolerate biomass hydrolysates. Here, we examined the furan derivatives tolerance in a novel and nonpathogenic species (strain BJa5) and in KT2440 using tolerance adaptive laboratory evolution (TALE) to enhance growth performance in a synthetic straw sugar cane hydrolysate enriched with furfural and 5-hydroxymethylfurfural (5-HMF). Initially, wild-type strains showed prolonged lag phases and low tolerance in the synthetic hydrolysate, but tolerance was improved after 90 days of sequential batch growth. Post-TALE, BJa5 and KT2440 end strains grew in synthetic hydrolysate containing 2 g/L furfural and 1 g/L 5-HMF at 48 and 24 h, respectively. Moreover, the KT2440 end strain notably grew in 2 g/L furfural and ≥1.7 g/L 5-HMF. Genome sequencing of end strains revealed mutations in genes and intergenic regions associated with transcriptional factors, acetate metabolism enzymes, environmental response proteins, and transposases. In a proof-of-concept experiment, the BJa5 end strain demonstrated the potential to detoxify synthetic hydrolysate by reducing the titers of acetate and furfural. This ability could enable industrial microorganisms, which are typically nontolerant to toxic hydrolysates, to be used for producing value-added compounds from biodetoxified hydrolysates.
包括KT2440在内的几种菌株具有广泛的代谢能力，可以同化木质素衍生化合物等生物质单体，但难以耐受生物质水解产物。在此，我们使用耐受性适应性实验室进化（TALE）方法，检测了一种新型非致病菌株（BJa5菌株）和KT2440对呋喃衍生物的耐受性，以提高其在富含糠醛和5-羟甲基糠醛（5-HMF）的合成甘蔗秸秆水解产物中的生长性能。最初，野生型菌株在合成水解产物中表现出延长的延迟期和低耐受性，但经过90天的连续分批培养后耐受性得到改善。TALE处理后，BJa5和KT2440的最终菌株分别在含有2 g/L糠醛和1 g/L 5-HMF的合成水解产物中于48小时和24小时内生长。此外，KT2440最终菌株在2 g/L糠醛和≥1.7 g/L 5-HMF中显著生长。最终菌株的基因组测序揭示了与转录因子、乙酸代谢酶、环境响应蛋白和转座酶相关的基因和基因间区域的突变。在一个概念验证实验中，BJa5最终菌株展示了通过降低乙酸和糠醛的滴度来解毒合成水解产物的潜力。这种能力可以使通常对有毒水解产物不耐受的工业微生物用于从生物解毒的水解产物中生产增值化合物。
