Table 1. Mutations found in the 156 T strain.

Locus_tag	Genome position	Timeline of the mutation	Mutation typea	Codon change	Description
TON_0536–0541	490749–496535	after 12th transfer	5786-bp deletion	frame shift	cytosolic NiFe-hydrogenase, formate transporter, formate dehydrogenase, iron-sulfur binding proteins
TON_0820	760634	after 12th transfer	substitution	Leu to Arg	aromatic amino acid permease
TON_1525	1400830	after 12th transfer	substitution	Thr to Ile	putative transcriptional regulator
TON_1544	1420154	after 62nd transfer	substitution	Pro to Leu	membrane protein
TON_0982	906462	after 32nd transfer	T insertion	frame shift	aminotransferase
TON_1548	1422235	after 102nd transfer	T deletion	frame shift	hypothetical protein
TON_1664	1527982	after 156th transfer	A deletion	frame shift	cation transporter
TON_1694	1559247	after 102nd transfer	12-bp insertion	frame shift	membrane-associated metalloprotease
TON_0544	499427–499765	after 156th transfer	338-bp deletion	frame shift	alcohol dehydrogenase
TON_1475–1476	1351559–1351609	after 62nd transfer	51-bp deletion	–	intergenic region

^aAll mutations were confirmed by PCR verification and Sanger sequencing.