Nā huahana

ʻO ka hoʻomaopopo ʻana i ka metabolism o ka microbial host he mea nui ia no ka hoʻomohala ʻana a me ka hoʻomohala ʻana i nā kaʻina biocatalytic e pili ana i ke kelepona holoʻokoʻa, no ka mea e kuhikuhi ana i ka pono hana.He ʻoiaʻiʻo kēia no ka biocatalysis redox kahi i hoʻohana ʻia ai nā cell metabolically active ma muli o ka cofactor/cosubstrate regenerative capacity endogenous i ka host.Ua hoʻohana ʻia ʻo Recombinant Escherichia coli no ka hana nui ʻana i ka proline-4-hydroxylase (P4H), he dioxygenase e hoʻopaʻa ana i ka hydroxylation o ka L-proline manuahi i loko o ka trans-4-hydroxy-L-proline me ka a-ketoglutarate (a-KG) ma ke ʻano he cosubstrate.I loko o kēia biocatalyst cell holoʻokoʻa, hāʻawi ke kikowaena kalapona kalapona i ka cosubstrate a-KG e pono ai, e hoʻopili pololei ana i ka hana biocatalytic P4H i ka metabolism kalapona a me ka hana metabolic.Ma ka hoʻohana ʻana i nā mea hana biology hoʻokolohua a me ka computational, e like me ka metabolic engineering a me (13) C-metabolic flux analysis ((13)C-MFA), ua noiʻi mākou a wehewehe nui i ka pane physiological, metabolic, a me ka bioenergetic o ka biocatalyst cell holoʻokoʻa. i ka bioconversion i manaʻo ʻia a ʻike ʻia nā bottlenecks metabolic hiki ke noʻonoʻo hou aku i ka ʻenekinia ala ala.ʻAʻole i alakaʻi ʻia nā biotransformations holoʻokoʻa holoʻokoʻa me kēia ʻano mutant i ka quantitative proline hydroxylation akā i ka pālua ʻana o ka helu hoʻokumu kikoʻī trans-4-L-hydroxyproline (hyp), i hoʻohālikelike ʻia me ka ʻano hihiu.Ua hōʻike ʻia ka ʻike ʻana o ka carbon flux ma waena o ka metabolism o ka mutant strain i ka piʻi ʻana o ke koi a-KG no ka hana P4H ʻaʻole i hoʻonui i ka flux hana a-KG, e hōʻike ana i kahi hana pōʻai TCA i hoʻoponopono paʻa ʻia ma lalo o nā kūlana i aʻo ʻia.Ma ka ʻano ʻano hihiu, P4H synthesis a me ka catalysis i hoʻemi i ka hua biomass.ʻO ka mea e mahalo ai, ua uku hou ka ΔputA i ka poho ATP a me NADH e pili ana ma o ka hoʻemi ʻana i nā koi o ka ikehu mālama ʻana i ka haʻahaʻa haʻahaʻa o ka glucose uptake rates, ma kahi o ka hoʻonui ʻana i ka hana TCA. e hoʻohiki no ka P4H catalysis huahua ʻaʻole wale i nā huaʻōlelo o ka biotransformation yield, akā e pili ana hoʻi i nā kumukūʻai no ka biotransformation a me ka uptake proline a me ka hua o ka hyp ma ke kumu ikehu.Hōʻike nā hualoaʻa, ma luna o kahi knockout putA, ʻo ka hui ʻana o ka TCA-cycle i proline hydroxylation ma o ka cosubstrate a-KG e lilo i kumu koʻikoʻi a me kahi pahuhopu e hoʻomaikaʻi hou i ka pono o nā biotransformations hilinaʻi a-KG.