Supplementary Fig. 1 Relative concentrations of amino acids after transamination reaction catalyzed by PpACL1, α- ketoglutarate as the amino acceptor. In the reaction, 18 different natural amino acids were used as amino donors. Control: AtASP2 purified protein was used as the positive control, empty vector pET22b or pET28a was used as a negative control, and H 2 O was also used as negative control instead of purified proteins. NH 4 + Substrate relative concentration (%) Substrate relative concentration (%)

NH 4 + Substrate relative concentration (%) NH 4 + Supplementary Fig. 2 Relative concentrations of amino acids after transamination reaction catalyzed by PpACL1, pyruvic acid as the amino acceptor. In the reaction, 18 different natural amino acids were used as amino donors. Control: AtASP2 purified protein was used as the positive control, empty vector pET22b or pET28a was used as a negative control, and H 2 O was also used as negative control instead of purified proteins.

NH 4 + Supplementary Fig. 3 Relative concentrations of amino acids after transamination reaction catalyzed by PpACL1, glyoxylic acid as the amino acceptor. In the reaction, 18 different natural amino acids were used as amino donors. Control: AtASP2 purified protein was used as the positive control, empty vector pET22b or pET28a was used as a negative control, and H 2 O was also used as negative control instead of purified proteins.

C D His-ACL1 I A pET28a I A His-ACL1 I A pET28a I A A His-ACL1 I A pET28a I A His-ACL1 I A pET28a I A B Supplementary Fig. 4 Detection of pyruvic acid after C-S lyase reaction catalyzed by PpACL1. In the reaction, Cystine (A), Cysteine (B), Cystathionine (C) or S-methyl-L-cysteine (D) were used as substrates respectively. I, inactivated enzyme, heated at 100℃ for 10min; A, activated enzyme.

pET28a I A His-ACL1 I A His-ACL2 I A pET28a I A His-ACL1 I A His-ACL2 I A A C B D pET28a I A His-ACL2 I A pET28a I A His-ACL2 I A E F Supplementary Fig. 5 Detection of pyruvic acid after C-S lyase reaction catalyzed by PpACL2. In the reaction, Cystine (A, B), Cysteine (C, D), Cystathionine (E) or S-methyl-L-cysteine (F) were used as substrates respectively. I, inactivated enzyme, heated at 100℃ for 10min; A, activated enzyme. Error bars show standard error (n=9).

— Negative control — pET28a — His-PpACL2 — AtASP2-His Asp Thr Ser Asn Gly Gln Ala Val Met Ile Leu Tyr Phe His Trp Lys NH4 + Arg Pro Glu Supplementary Fig. 6 Chromatogram of amino acids after transamination reactions catalyzed by PpACL2. In the reaction, α- ketoglutarate was used as the amino acceptor, 18 different natural amino acids were used as amino donors. Control: AtASP2 purified protein was used as the positive control, empty vector pET28a was used as a negative control, and H 2 O was also used as negative control instead of purified proteins. L-glutamic acid was marked with red arrow Detector response (mv) Retention time (min)