{"id":{"repo_id":"nus","oai_identifier":"oai:scholarbank.nus.edu.sg:10635/239378"},"canonical_url":"https://search.dev.ndltd.org/etd/nus/oai:scholarbank.nus.edu.sg:10635/239378","repository":{"repo_id":"nus","name":"National University of Singapore","base_url":"https://scholarbank.nus.edu.sg/oai/request"},"display":{"title":"ENZYME ENGINEERING AND GREEN SYNTHESIS OF N-ACYL GLYCINE BIOSURFACTANTS FROM RENEWABLE RAW MATERIALS","abstract":"Amide syntheses remain a key challenging green chemistry reaction. For instance, green synthesis of N-acyl glycines as biosurfactants and therapeutics is highly desirable to replace chemical pathways using toxic phosgene. Herein, we report a novel concept for enzymatic amidation in an aqueous system via glycerol activation of fatty acids and their subsequent aminolysis with glycine to synthesize N-acyl glycines. We then engineer an enzyme (proRML) by reshaping its catalytic pocket to enhance its aminolysis activity and catalytic efficiency by 103-fold and 465-fold, respectively. The evolved proRML (D156S/L258K/L267N/S83D/L58K/R86K/W88V) catalyzed the amidation of a fatty acid with glycine to give N-lauroylglycine with high yield (80 %). It accepts a broad range of medium- to long-chain fatty acids (C8–C18), giving high yields of N-decanoyl-, N-myristoyl-, and N-oleoylglycine. The developed amidation concept may be general, and the engineered enzyme is useful for the green synthesis of N-acyl glycines.","abstract_html":"Amide syntheses remain a key challenging green chemistry reaction. For instance, green synthesis of N-acyl glycines as biosurfactants and therapeutics is highly desirable to replace chemical pathways using toxic phosgene. Herein, we report a novel concept for enzymatic amidation in an aqueous system via glycerol activation of fatty acids and their subsequent aminolysis with glycine to synthesize N-acyl glycines. We then engineer an enzyme (proRML) by reshaping its catalytic pocket to enhance its aminolysis activity and catalytic efficiency by 103-fold and 465-fold, respectively. The evolved proRML (D156S/L258K/L267N/S83D/L58K/R86K/W88V) catalyzed the amidation of a fatty acid with glycine to give N-lauroylglycine with high yield (80 %). It accepts a broad range of medium- to long-chain fatty acids (C8–C18), giving high yields of N-decanoyl-, N-myristoyl-, and N-oleoylglycine. The developed amidation concept may be general, and the engineered enzyme is useful for the green synthesis of N-acyl glycines.","abstract_has_math":false,"creators":["GLEN KUA KAI BIN"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2023,"date_issued":"2023-01-20","date_published":"2023-01-20","updated_at":"2026-07-24T03:30:34Z","subjects":["Protein Engineering","N-acylglycines","Ester-Amide Interconversion","Directed Evolution","Aminolysis","Amide Synthesis"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":null,"outbound_label":null,"outbound_source":null},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:creator","label":"Author","values":["GLEN KUA KAI BIN"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.issued","label":"Date","values":["2023-01-20"]},{"key":"dc:relation.isreferencedby","label":"Dc Relation Isreferencedby","values":["https://scholarbank.nus.edu.sg/handle/10635/239378"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Protein Engineering","N-acylglycines","Ester-Amide Interconversion","Directed Evolution","Aminolysis","Amide Synthesis"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://scholarbank.nus.edu.sg/bitstreams/bd6d1ca7-51c2-458b-94ef-300aa2b69177/download"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Amide syntheses remain a key challenging green chemistry reaction. For instance, green synthesis of N-acyl glycines as biosurfactants and therapeutics is highly desirable to replace chemical pathways using toxic phosgene. Herein, we report a novel concept for enzymatic amidation in an aqueous system via glycerol activation of fatty acids and their subsequent aminolysis with glycine to synthesize N-acyl glycines. We then engineer an enzyme (proRML) by reshaping its catalytic pocket to enhance its aminolysis activity and catalytic efficiency by 103-fold and 465-fold, respectively. The evolved proRML (D156S/L258K/L267N/S83D/L58K/R86K/W88V) catalyzed the amidation of a fatty acid with glycine to give N-lauroylglycine with high yield (80 %). It accepts a broad range of medium- to long-chain fatty acids (C8–C18), giving high yields of N-decanoyl-, N-myristoyl-, and N-oleoylglycine. The developed amidation concept may be general, and the engineered enzyme is useful for the green synthesis of N-acyl glycines."]},{"key":"dc:format.checksum.md5","label":"Dc Format Checksum Md5","values":["d247982464599ecb31c96665ee43fbf7","4f83c01ddef6e7894315e163b9deb2ad"]},{"key":"dc:title","label":"Title","values":["ENZYME ENGINEERING AND GREEN SYNTHESIS OF N-ACYL GLYCINE BIOSURFACTANTS FROM RENEWABLE RAW MATERIALS"]}]}],"canonical_facts":{"dc:creator":["GLEN KUA KAI BIN"],"dc:date.issued":["2023-01-20"],"dc:description.abstract":["Amide syntheses remain a key challenging green chemistry reaction. For instance, green synthesis of N-acyl glycines as biosurfactants and therapeutics is highly desirable to replace chemical pathways using toxic phosgene. Herein, we report a novel concept for enzymatic amidation in an aqueous system via glycerol activation of fatty acids and their subsequent aminolysis with glycine to synthesize N-acyl glycines. We then engineer an enzyme (proRML) by reshaping its catalytic pocket to enhance its aminolysis activity and catalytic efficiency by 103-fold and 465-fold, respectively. The evolved proRML (D156S/L258K/L267N/S83D/L58K/R86K/W88V) catalyzed the amidation of a fatty acid with glycine to give N-lauroylglycine with high yield (80 %). It accepts a broad range of medium- to long-chain fatty acids (C8–C18), giving high yields of N-decanoyl-, N-myristoyl-, and N-oleoylglycine. The developed amidation concept may be general, and the engineered enzyme is useful for the green synthesis of N-acyl glycines."],"dc:format.checksum.md5":["d247982464599ecb31c96665ee43fbf7","4f83c01ddef6e7894315e163b9deb2ad"],"dc:identifier.uri":["https://scholarbank.nus.edu.sg/bitstreams/bd6d1ca7-51c2-458b-94ef-300aa2b69177/download"],"dc:relation.isreferencedby":["https://scholarbank.nus.edu.sg/handle/10635/239378"],"dc:subject":["Protein Engineering","N-acylglycines","Ester-Amide Interconversion","Directed Evolution","Aminolysis","Amide Synthesis"],"dc:title":["ENZYME ENGINEERING AND GREEN SYNTHESIS OF N-ACYL GLYCINE BIOSURFACTANTS FROM RENEWABLE RAW MATERIALS"],"dc:type":["Thesis"]},"updated_at":"2026-07-24T03:30:34Z"}