{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,11]],"date-time":"2026-05-11T01:20:29Z","timestamp":1778462429233,"version":"3.51.4"},"reference-count":30,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T00:00:00Z","timestamp":1706745600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T00:00:00Z","timestamp":1706745600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100022963","name":"Key Research and Development Program of Zhejiang Province","doi-asserted-by":"publisher","award":["2021C03088"],"award-info":[{"award-number":["2021C03088"]}],"id":[{"id":"10.13039\/100022963","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Bioresour. Bioprocess."],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Biliverdin, a bile pigment hydrolyzed from heme by heme oxygenase (HO), serves multiple functions in the human body, including antioxidant, anti-inflammatory, and immune response inhibitory activities. Biliverdin has great potential as a clinical drug; however, no economic and efficient production method is available currently. Therefore, the production of biliverdin by the biotransformation of exogenous heme using recombinant HO-expressing yeast cells was studied in this research. First, the heme oxygenase-1 gene (<jats:italic>HO1<\/jats:italic>) encoding the inducible plastidic isozyme from <jats:italic>Arabidopsis thaliana<\/jats:italic>, with the plastid transport peptide sequence removed, was recombined into <jats:italic>Pichia pastoris<\/jats:italic> GS115 cells. This resulted in the construction of a recombinant <jats:italic>P. pastoris<\/jats:italic> GS115-HO1 strain that expressed active HO1 in the cytoplasm. After that, the concentration of the inducer methanol, the induction culture time, the pH of the medium, and the concentration of sorbitol supplied in the medium were optimized, resulting in a significant improvement in the yield of HO1. Subsequently, the whole cells of GS115-HO1 were employed as catalysts to convert heme chloride (hemin) into biliverdin. The results showed that the yield of biliverdin was 132\u00a0mg\/L when hemin was added to the culture of GS115-HO1 and incubated for 4\u00a0h at 30\u00a0\u00b0C. The findings of this study have laid a good foundation for future applications of this method for the economical production of biliverdin.<\/jats:p>\n                <jats:p><jats:bold>Graphical Abstract<\/jats:bold><\/jats:p>","DOI":"10.1186\/s40643-024-00736-w","type":"journal-article","created":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T16:02:12Z","timestamp":1706803332000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Production of biliverdin by biotransformation of exogenous heme using recombinant Pichia pastoris cells"],"prefix":"10.1186","volume":"11","author":[{"given":"Jianfeng","family":"Mei","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yanchao","family":"Han","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shihang","family":"Zhuang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhikai","family":"Yang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu","family":"Yi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5683-7561","authenticated-orcid":false,"given":"Guoqing","family":"Ying","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,2,1]]},"reference":[{"issue":"6","key":"736_CR1","doi-asserted-by":"publisher","DOI":"10.1242\/jeb.200055","volume":"222","author":"JL Baylor","year":"2019","unstructured":"Baylor JL, Butler MW (2019) Immune challenge-induced oxidative damage may be mitigated by biliverdin. J Exp Biol 222(6):jeb200055. https:\/\/doi.org\/10.1242\/jeb.200055","journal-title":"J Exp Biol"},{"key":"736_CR2","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1016\/j.sbi.2019.01.006","volume":"59","author":"AI Celis","year":"2019","unstructured":"Celis AI, Dubois JL (2019) Making and breaking heme. Curr Opin Struct Biol 59:19\u201328. https:\/\/doi.org\/10.1016\/j.sbi.2019.01.006","journal-title":"Curr Opin Struct Biol"},{"key":"736_CR3","doi-asserted-by":"publisher","first-page":"89","DOI":"10.1186\/1472-6750-12-89","volume":"12","author":"D Chen","year":"2012","unstructured":"Chen D, Brown JD, Kawasaki Y, Bommer J, Takemoto JY (2012) Scalable production of biliverdin IX\u03b1 by Escherichia coli. BMC Biotechnol 12:89. https:\/\/doi.org\/10.1186\/1472-6750-12-89","journal-title":"BMC Biotechnol"},{"issue":"6","key":"736_CR4","doi-asserted-by":"publisher","first-page":"2050","DOI":"10.13344\/j.microbiol.china.210944","volume":"49","author":"LJ Chen","year":"2022","unstructured":"Chen LJ, Xu CL, Su Y, Xiao GN, Xu ZN, Wei PL (2022) Optimization of leghemoglobin expression conditions in Pichia pastoris (in Chinese). Microbiol China 49(6):2050\u20132061. https:\/\/doi.org\/10.13344\/j.microbiol.china.210944","journal-title":"Microbiol China"},{"issue":"8","key":"736_CR5","doi-asserted-by":"publisher","first-page":"927","DOI":"10.1023\/a:1019974822667","volume":"67","author":"ZK Ding","year":"2002","unstructured":"Ding ZK, Xu YQ (2002) Purification and characterization of biliverdin IX\u03b1 from Atlantic salmon (Salmo salar) bile. Biochem Mosc 67(8):927\u2013932. https:\/\/doi.org\/10.1023\/a:1019974822667","journal-title":"Biochem Mosc"},{"issue":"1","key":"736_CR6","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1016\/0305-0491(90)90174-r","volume":"97","author":"LS Fang","year":"1990","unstructured":"Fang LS, Bada JL (1990) The blue-green blood plasma of marine fish. Comp Biochem Physiol Part B Biochem Mol Biol 97(1):37\u201345. https:\/\/doi.org\/10.1016\/0305-0491(90)90174-r","journal-title":"Comp Biochem Physiol Part B Biochem Mol Biol"},{"issue":"7","key":"736_CR7","doi-asserted-by":"publisher","first-page":"706","DOI":"10.1038\/nchembio.2390","volume":"13","author":"J Fernandez-Rodriguez","year":"2017","unstructured":"Fernandez-Rodriguez J, Moser F, Song M, Voigt CA (2017) Engineering RGB color vision into Escherichia coli. Nat Chem Biol 13(7):706\u2013708. https:\/\/doi.org\/10.1038\/nchembio.2390","journal-title":"Nat Chem Biol"},{"key":"736_CR8","doi-asserted-by":"publisher","first-page":"466","DOI":"10.1016\/j.freeradbiomed.2016.06.016","volume":"97","author":"E Gonzalez-Sanchez","year":"2016","unstructured":"Gonzalez-Sanchez E, Perez MJ, Nytofte NS, Briz O, Monte MJ, Lozano E, Serrano MA, Marin JJG (2016) Protective role of biliverdin against bile acid-induced oxidative stress in liver cells. Free Radical Biol Med 97:466\u2013477. https:\/\/doi.org\/10.1016\/j.freeradbiomed.2016.06.016","journal-title":"Free Radical Biol Med"},{"issue":"10","key":"736_CR9","doi-asserted-by":"publisher","first-page":"1533","DOI":"10.1021\/acs.jchemed.7b00449","volume":"94","author":"S Halepas","year":"2017","unstructured":"Halepas S, Hamchand R, Lindeyer SED, Br\u00fcckner C (2017) Isolation of biliverdin ix\u03b1, as its dimethyl ester, from emu eggshells. J Chem Educ 94(10):1533\u20131537. https:\/\/doi.org\/10.1021\/acs.jchemed.7b00449","journal-title":"J Chem Educ"},{"issue":"3","key":"736_CR10","doi-asserted-by":"publisher","first-page":"505","DOI":"10.1007\/s00360-018-1146-5","volume":"188","author":"MP Homsher","year":"2018","unstructured":"Homsher MP, Astor MT, Hines JK, Butler MW (2018) Immune challenges decrease biliverdin concentration in the spleen of northern Bobwhite quail. Colinus Virginianus J Comp Physiol B 188(3):505\u2013515. https:\/\/doi.org\/10.1007\/s00360-018-1146-5","journal-title":"Colinus Virginianus J Comp Physiol B"},{"issue":"1","key":"736_CR11","doi-asserted-by":"publisher","first-page":"161","DOI":"10.1111\/j.1432-1033.1991.tb16357.x","volume":"202","author":"K Ishikawa","year":"1991","unstructured":"Ishikawa K, Sato M, Yoshida T (1991) Expression of rat heme oxygenase in Escherichia coli as a catalytically active, full-length form that binds to bacterial membranes. Eur J Biochem 202(1):161\u2013165. https:\/\/doi.org\/10.1111\/j.1432-1033.1991.tb16357.x","journal-title":"Eur J Biochem"},{"key":"736_CR12","doi-asserted-by":"publisher","first-page":"1165","DOI":"10.2147\/ndt.s300773","volume":"17","author":"J Li","year":"2021","unstructured":"Li J, Peng L, Bai W, Peng P, Chen W, Yang W, Shao J (2021) Biliverdin protects against cerebral ischemia\/reperfusion injury by regulating the miR-27a-3p\/Rgs1 axis. Neuropsychiatr Dis Treat 17:1165\u20131181. https:\/\/doi.org\/10.2147\/ndt.s300773","journal-title":"Neuropsychiatr Dis Treat"},{"issue":"3","key":"736_CR13","doi-asserted-by":"publisher","first-page":"198","DOI":"10.1038\/84915","volume":"8","author":"AF Mcdonagh","year":"2001","unstructured":"Mcdonagh AF (2001) Turning green to gold. Nat Struct Biol 8(3):198\u2013200. https:\/\/doi.org\/10.1038\/84915","journal-title":"Nat Struct Biol"},{"issue":"5","key":"736_CR14","doi-asserted-by":"publisher","first-page":"814","DOI":"10.1016\/j.freeradbiomed.2010.06.001","volume":"49","author":"AF Mcdonagh","year":"2010","unstructured":"Mcdonagh AF (2010) The biliverdin-bilirubin antioxidant cycle of cellular protection: missing a wheel? Free Radic Biol Med 49(5):814\u2013820. https:\/\/doi.org\/10.1016\/j.freeradbiomed.2010.06.001","journal-title":"Free Radic Biol Med"},{"issue":"3","key":"736_CR15","doi-asserted-by":"publisher","first-page":"563","DOI":"10.1007\/s00449-021-02679-4","volume":"45","author":"J Mei","year":"2022","unstructured":"Mei J, Wu X, Zheng S, Yi Y, Wang X, Ying G (2022) Production of bilirubin by biotransformation of biliverdin using recombinant Escherichia coli cells. Bioprocess Biosyst Eng 45(3):563\u2013571. https:\/\/doi.org\/10.1007\/s00449-021-02679-4","journal-title":"Bioprocess Biosyst Eng"},{"issue":"2","key":"736_CR16","doi-asserted-by":"publisher","first-page":"394","DOI":"10.1016\/j.atherosclerosis.2012.07.043","volume":"224","author":"JA Moraes","year":"2012","unstructured":"Moraes JA, Barcellos-De-Souza P, Rodrigues G, Nascimento-Silva V, Silva SV, Assreuy J, Arruda MA, Barja-Fidalgo C (2012) Heme modulates smooth muscle cell proliferation and migration via NADPH oxidase: a counter-regulatory role for heme oxygenase system. Atherosclerosis 224(2):394\u2013400. https:\/\/doi.org\/10.1016\/j.atherosclerosis.2012.07.043","journal-title":"Atherosclerosis"},{"issue":"8","key":"736_CR17","doi-asserted-by":"publisher","DOI":"10.1002\/bies.202000010","volume":"42","author":"J Morales","year":"2020","unstructured":"Morales J (2020) Eggshell biliverdin as an antioxidant maternal effect: biliverdin as an antioxidant resource in oviparous animals. BioEssays 42(8):e2000010. https:\/\/doi.org\/10.1002\/bies.202000010","journal-title":"BioEssays"},{"issue":"4","key":"736_CR18","doi-asserted-by":"publisher","first-page":"587","DOI":"10.1161\/circulationaha.104.509778","volume":"112","author":"A Nakao","year":"2005","unstructured":"Nakao A, Murase N, Ho C, Toyokawa H, Billiar TR, Kanno S (2005) Biliverdin administration prevents the formation of intimal hyperplasia induced by vascular injury. Circulation 112(4):587\u2013591. https:\/\/doi.org\/10.1161\/circulationaha.104.509778","journal-title":"Circulation"},{"issue":"12","key":"736_CR19","doi-asserted-by":"publisher","first-page":"2217","DOI":"10.1002\/prot.25386","volume":"85","author":"Y Naoe","year":"2017","unstructured":"Naoe Y, Nakamura N, Rahman MM, Tosha T, Nagatoishi S, Tsumoto K, Shiro Y, Sugimoto H (2017) Structural basis for binding and transfer of heme in bacterial heme-acquisition systems. Proteins 85(12):2217\u20132230. https:\/\/doi.org\/10.1002\/prot.25386","journal-title":"Proteins"},{"key":"736_CR20","unstructured":"Pendrak ML, Roberts DD (2005) Methods for the production of biliverdin. US 20050209305, 2005-09-22. https:\/\/image-ppubs.uspto.gov\/dirsearch-public\/print\/downloadPdf\/20050209305"},{"issue":"1","key":"736_CR21","doi-asserted-by":"publisher","first-page":"28","DOI":"10.1186\/s13036-019-0156-5","volume":"13","author":"J Seok","year":"2019","unstructured":"Seok J, Ko YJ, Lee M-E, Hyeon JE, Han SO (2019) Systems metabolic engineering of Corynebacterium glutamicum for the bioproduction of biliverdin via protoporphyrin independent pathway. J Biol Eng 13(1):28. https:\/\/doi.org\/10.1186\/s13036-019-0156-5","journal-title":"J Biol Eng"},{"key":"736_CR22","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejps.2020.105546","volume":"155","author":"RG Shiels","year":"2020","unstructured":"Shiels RG, Hewage W, Pennell EN, Vidimce J, Grant G, Pearson AG, Wagner KH, Morgan M, Bulmer AC (2020) Biliverdin and bilirubin sulfonate inhibit monosodium urate induced sterile inflammation in the rat. Eur J Pharm Sci 155:105546. https:\/\/doi.org\/10.1016\/j.ejps.2020.105546","journal-title":"Eur J Pharm Sci"},{"issue":"17","key":"736_CR23","doi-asserted-by":"publisher","first-page":"2964","DOI":"10.1016\/j.febslet.2014.05.050","volume":"588","author":"AY Shin","year":"2014","unstructured":"Shin AY, Han YJ, Song PS, Kim JI (2014) Expression of recombinant full-length plant phytochromes assembled with phytochromobilin in Pichia pastoris. FEBS Lett 588(17):2964\u20132970. https:\/\/doi.org\/10.1016\/j.febslet.2014.05.050","journal-title":"FEBS Lett"},{"issue":"45","key":"736_CR24","doi-asserted-by":"publisher","first-page":"11962","DOI":"10.1073\/pnas.1707190114","volume":"114","author":"Y Uda","year":"2017","unstructured":"Uda Y, Goto Y, Oda S, Kohchi T, Matsuda M, Aoki K (2017) Efficient synthesis of phycocyanobilin in mammalian cells for optogenetic control of cell signaling. Proc Natl Acad Sci USA 114(45):11962\u201311967. https:\/\/doi.org\/10.1073\/pnas.1707190114","journal-title":"Proc Natl Acad Sci USA"},{"issue":"10","key":"736_CR25","doi-asserted-by":"publisher","first-page":"1450","DOI":"10.1016\/j.chembiol.2019.07.006","volume":"26","author":"C Vasavda","year":"2019","unstructured":"Vasavda C, Kothari R, Malla AP, Tokhunts R, Lin A, Ji M, Ricco C, Xu R, Saavedra HG, Sbodio JI, Snowman AM, Albacarys L, Hester L, Sedlak TW, Paul BD, Snyder SH (2019) Bilirubin links heme metabolism to neuroprotection by scavenging superoxide. Cell Chem Biol 26(10):1450\u20131460. https:\/\/doi.org\/10.1016\/j.chembiol.2019.07.006","journal-title":"Cell Chem Biol"},{"issue":"13","key":"736_CR26","doi-asserted-by":"publisher","first-page":"6146","DOI":"10.1039\/c3cp54307b","volume":"16","author":"T Watermann","year":"2014","unstructured":"Watermann T, Elgabarty H, Sebastiani D (2014) Phycocyanobilin in solution \u2013 a solvent triggered molecular switch. Phys Chem Chem Phys 16(13):6146\u20136152. https:\/\/doi.org\/10.1039\/c3cp54307b","journal-title":"Phys Chem Chem Phys"},{"issue":"30","key":"736_CR27","doi-asserted-by":"publisher","first-page":"22357","DOI":"10.1016\/S0021-9258(18)41536-0","volume":"268","author":"A Wilks","year":"1993","unstructured":"Wilks A, Ortiz De Montellano PR (1993) Rat liver heme oxygenase. High level expression of a truncated soluble form and nature of the meso-hydroxylating species. J Bio Chem 268(30):22357\u201322362. https:\/\/doi.org\/10.1016\/S0021-9258(18)41536-0","journal-title":"J Bio Chem"},{"issue":"7","key":"736_CR28","doi-asserted-by":"publisher","first-page":"2581","DOI":"10.13345\/j.cjb.220137","volume":"38","author":"S Yan","year":"2022","unstructured":"Yan S, Shao M, Xu M, Zhang X, Yang T, Rao Z (2022) Efficient production of biliverdin through whole-cell biocatalysis using recombinant Escherichia coli. Chin J Biotechnol 38(7):2581\u20132593. https:\/\/doi.org\/10.13345\/j.cjb.220137","journal-title":"Chin J Biotechnol"},{"issue":"3","key":"736_CR29","doi-asserted-by":"publisher","first-page":"546","DOI":"10.1093\/ps\/85.3.546","volume":"85","author":"R Zhao","year":"2006","unstructured":"Zhao R, Xu GY, Liu ZZ, Li JY, Yang N (2006) A study on eggshell pigmentation: biliverdin in blue-shelled chickens. Poult Sci 85(3):546\u2013549. https:\/\/doi.org\/10.1093\/ps\/85.3.546","journal-title":"Poult Sci"},{"issue":"8","key":"736_CR30","doi-asserted-by":"publisher","first-page":"2116","DOI":"10.1038\/bjc.2014.98","volume":"110","author":"J Zheng","year":"2014","unstructured":"Zheng J, Nagda DA, Lajud SA, Kumar S, Mouchli A, Bezpalko O, Omalley BW Jr, Li D (2014) Biliverdin\u2019s regulation of reactive oxygen species signalling leads to potent inhibition of proliferative and angiogenic pathways in head and neck cancer. Br J Cancer 110(8):2116\u20132122. https:\/\/doi.org\/10.1038\/bjc.2014.98","journal-title":"Br J Cancer"}],"container-title":["Bioresources and Bioprocessing"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s40643-024-00736-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s40643-024-00736-w\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s40643-024-00736-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T16:02:43Z","timestamp":1706803363000},"score":1,"resource":{"primary":{"URL":"https:\/\/bioresourcesbioprocessing.springeropen.com\/articles\/10.1186\/s40643-024-00736-w"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,2,1]]},"references-count":30,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,12]]}},"alternative-id":["736"],"URL":"https:\/\/doi.org\/10.1186\/s40643-024-00736-w","relation":{},"ISSN":["2197-4365"],"issn-type":[{"value":"2197-4365","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,2,1]]},"assertion":[{"value":"3 December 2023","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"22 January 2024","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"1 February 2024","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"This study did not involve either human participants, human data and human tissue or animals. As such additional consent for publication beyond that of each author is not required.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Each author has consented to the publication of this study, incl. main manuscript and supporting information.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"There are no competing interests regarding this study.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"19"}}