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. 2021 Apr 15;105(9):3423–3456. doi: 10.1007/s00253-021-11269-z

Table 2.

Use of the pBBR1MCS-based plasmid family in Gluconobacter. The studies are grouped by the pBBR1MCS derivative and ordered by the year of publication. cyt. cytochrome; DH dehydrogenase; DHA dihydroxyacetone; GFP green fluorescent protein; P promoter; PPP pentose phosphate pathway; PQQ pyrroloquinoline quinone; SNP single nucleotide polymorphism; SP signal peptide

Expressed genes Encoded activity/function P a Context Reference
pBBR1MCS-5
tldD, pqqA, pqqABCDE from G. oxydans 621H PQQ biosynthesis-related enzymes PtufB, analysis of PQQ biosynthesis genes in G. oxydans Hölscher and Görisch 2006
PpqqA,
PpqqBCDE
gdh, ga5dh from G. oxydans 621H membrane-bound PQQ-dependent glucose DH and gluconate-5-DH Plac increase of gluconate and 5-keto-d-gluconic acid production by G. oxydans strains Merfort et al. 2006b
ga5dh from G. oxydans 621H membrane-bound PQQ-dependent gluconate-5-DH PtufB, increase of 5-keto-d-gluconic acid production by G. oxydans mutant strain Merfort et al. 2006a
Pgdh
sldAB from G. oxydans 621H membrane-bound PQQ-dependent polyol DH PtufB, increase of DHA production from glycerol by G. oxydans mutant Gätgens et al. 2007
Pgdh
sldAB from G. oxydans M5 membrane-bound PQQ-dependent d-sorbitol DH Plac characterization of the enzyme in G. oxydans M5 and role in miglitol precursor production Yang et al. 2008
sldBA from G. suboxydans IFO 3255 membrane-bound PQQ-dependent d-sorbitol DH Plac, characterization of two membrane-bound d-sorbitol DHs and complementation in G. frateurii Soemphol et al. 2008
PsldBA
sldA’B’ from G. oxydans 621H N-terminal fragments of membrane-bound PQQ-dependent polyol DH PtufB construction of the pEXGOX family; a cloning and expression vector system with PtufB from G. oxydans Schleyer et al. 2008
adhAB from G. oxydans DSM 2003 membrane-bound PQQ-dependent alcohol DH (mADH) Plac enzyme characterization; mADH mutant complementation; lactic acid production from 1,2-propanediol Wei et al. 2010
gdh from G. oxydans M5 membrane-bound PQQ-dependent glycerol DH Plac enhanced production of DHA by G. oxydans mADH mutant Li et al. 2010b
vgb from Vitreoscilla Vitreoscilla hemoglobin PtufB improvement of cell growth and DHA production by G. oxydans M5 Li et al. 2010a
adhAB from G. oxydans DSM 2003 membrane-bound PQQ-dependent alcohol DH (mADH) Padh characterization of mADH and complementation in G. oxydans DSM 2003 mADH mutant Wei et al. 2012
xdh from G. oxydans NH-10; gdh from B. subtilis xylitol DH (XDH) and glucose DH (GDH) PtufB improvement of xylitol production by construction of a xdh and gdh co-expressing G. oxydans strain Zhang et al. 2013
ndh and ga2dh from G. oxydans DSM 2003; gfp membrane-bound type II NADH DH; FAD-dependent gluconate-2-DH; GFP PgHp0169, evaluation of promoter gHp0169 and improvement of 2-keto-d-gluconic acid production by G. oxydans DSM 2003 Shi et al. 2014
PtufB
ga2dh from G. oxydans DSM 2003 FAD-dependent gluconate-2-DH PtufB rational mutagenesis to increase the pBBR1MCS-5 copy number; 2-keto-d-gluconic acid production Shi et al. 2015
sldAB from G. oxydans 621H; gfp membrane-bound PQQ-dependent polyol DH; green fluorescent protein PGOX0169, PGOX0264, PGOX0452, PtufB promoter strength analysis and 5-keto-d-gluconate accumulation using combinatorial metabolic engineering strategies in industrial strain G. oxydans ZJU2 Yuan et al. 2016a
ga2dh from G. oxydans DSM 2003 FAD-dependent gluconate-2-DH PtufB, Pga2dh, PgHp0169 improvement of 2-keto-d-gluconic acid production by G. oxydans DSM 2003 Li et al. 2016a
adhS and adhABS from G. oxydans DSM 2003 membrane-bound PQQ-dependent alcohol DH (mADH) PadhS, PadhAB study of the AdhS subunit and effect on mADH activity in G. oxydans DSM 2003; hydroxy acid production Zhang et al. 2016
gcd from X. campestris DSM 3586 alcohol DH PGOX0169 improvement of 5-keto-d-gluconate production by G. oxydans ZJU2 mutant strain Yuan et al. 2016b
zwf, gnd from G. oxydans NH-10 glucose-6-phosphate DH and 6-phospho-gluconate DH PtufB improvement of xylitol production by G. oxydans NH-10; coenzyme regeneration efficiency of the PPP Li et al. 2016b
fusion of L-RI gene from Acinetobacter sp. DL-28; sdh from G. oxydans DSM 2003; gfp l-ribose isomerase (L-RI) C-terminally fused to PDZ ligand; membrane-bound PQQ-dependent d-sorbitol DH (SDH) N-terminally fused to PDZ domain; GFP fusion protein PtufB protein localization; novel strategy for the production of l-erythrose by localizing the assembly of L-RI to SDH via protein-peptide interaction of PDZ ligand and PDZ domain Zou et al. 2017
adh, idh, aldh, gidh, gdh, pqq3, pqq4, sldAB from G. oxydans 621H membrane-bound DHs (mDHs) PGOX1067-68, PGOX1857, test of promoter strength and enzymatic characterization of mDHs in G. oxydans multi-deletion strain BP.9 Mientus et al. 2017
genes of putative membrane-bound DHs from mother of vinegar DNA glucose DH like DH; alcohol DH like DH; polyol DH like DH; aldehyde DH like DH; d-lactate DH like DH Padh (PGOX1067-68) heterologous expression of metagenomic mDHs in G. oxydans BP.9 and identification of new substrate specificities Peters et al. 2017
adhA, adhAB, aldh from G. oxydans DSM 2003 membrane-bound alcohol DH (mADH) and aldehyde DH (mALDH) PtufB characterization of mADH and mALDH; 3-hydroxypropionic acid production Zhu et al. 2018
sldAB from G. oxydans ZJB-605 membrane-bound PQQ-dependent d-sorbitol DH PgHp0169 improvement of miglitol precursor production by industrial strain G. oxydans ZJB-605 Ke et al. 2019
trx from G. oxydans NL71 thioredoxin Plac enhancement of G. oxydans resistance to lignocellulosic-derived inhibitors; xylonic acid production Shen et al. 2020
araC, araE from E. coli MC4100 transcriptional regulator AraC; l-arabinose-transporter AraE ParaBAD plasmid-based tunable l-arabinose-inducible expression system for G. oxydans Fricke et al. 2020
pqqABCDE and tldD from G. oxydans H-8 PQQ biosynthesis-related enzymes PgHp0169 increase PQQ biosynthesis in repeated batch biotransformation for miglitol precursor production by mutant strain G. oxydans H-8 Liu et al. 2020a
pBBR1MCS-2
gdh from G. oxydans 621H membrane-bound PQQ-dependent glucose DH Plac gluconate and 5-keto-d-gluconic acid accumulation by G. oxydans Merfort et al. 2006b
GOX1857 from G. oxydans 621H membrane-bound PQQ-dependent inositol DH PGOX1857 substrate and co-factor analysis of GOX1857 in G. oxydans 621H Hölscher et al. 2007
uidA from E. coli DH5α β-d-glucuronidase UidA Plac, PGOX0254, PGOX0452 characterization of constitutive G. oxydans promoter strength Kallnik et al. 2010
sldA from G. oxydans DSM 7145 membrane-bound PQQ-dependent polyol / glycerol DH PsldB characterization of sldA disruptant mutant and substrate analysis of the enzyme Voss et al. 2010
mgdh from G. oxydans 621H membrane-bound PQQ-dependent glucose DH Pmgdh evolution for enhanced growth and biotransformations by G. oxydans at low glucose concentration Zhu et al. 2011
edd-eda and gnd from G. oxydans 621H 6-phosphogluconate dehydratase; 2-keto-3-deoxy-6-phospho-gluconate aldolase; 6-phosphogluconate DH PGOX0384 analysis of cytoplasmic fructose catabolism in a Δgnd mutant lacking the oxidative PPP and Δedd Δeda mutant lacking the Entner-Doudoroff pathway Richhardt et al. 2012
cyoBACD, cydAB and cydABCD of G. oxydans 621H quinol terminal oxidases cytochrome bo3 and cytochrome bd PGOX0384 influence of terminal oxidases on growth and yield of G. oxydans 621H Richhardt et al. 2013
GOX0265 of G. oxydans 621H membrane-bound PQQ-dependent glucose DH (mGHD) with C-terminal Strep-tag PGOX0452 gluconate production; influence of mGDH on O2 consumption rate; membrane protein purification by Strep-tactin affinity chromatography Meyer et al. 2013
phoA and treA from E. coli DH5α alkaline phosphatase and trehalase fused to selected SPs PGOX0264 SP screening in G. oxydans and broadening the substrate range of G. oxydans for growth on trehalose Kosciow et al. 2014
sldhAB from G. oxydans WSH-003 membrane-bound PQQ-dependent sorbitol DH (SLDH) PtufB enhancing production of l-sorbose from d-sorbitol; increase sldhAB mRNA level by artificial poly-A/T tail Xu et al. 2014
ndh from G. oxydans DSM 3504 type II NADH DH (NDH-2) Pndh consequence of an additional NADH DH paralog on growth of G. oxydans 621H Kostner et al. 2015
sdhCDAB, sdhE A. pasteurianus; sdhE from Serratia sp. succinate DH (SDH) and assembly factor SdhE for attachment of FAD to SdhA

PGOX0384,

PsdhE-Ace, PsdhE-Ser

SdhE-dependent formation of a functional A. pasteurianus SDH in G. oxydans 621H Kiefler et al. 2015
GOX1801 from G. oxydans 621H succinic semialdehyde reductase (SSR) fused C-terminally to Strep-tag PGOX0264 characterization of GOX1801 enzyme activities; the first SSR from an aerobic bacterium with high catalytic efficiency Meyer et al. 2015
sldh from G. oxydans WSH-003; egfp d-sorbitol DH; enhanced GFP (eGFP), PB932_2000, PtufB identification of strong promoters; comparison of PB932_2000 with PtufB form E. coli and from G. oxydans; enhancing production of l-sorbose Hu et al. 2015
GOX1432 from G. oxydans 621H cytoplasmic NADP+-dependent mannitol DH fused C-terminally to Strep-tag II PGOX0264 role of mannitol DHs in osmoprotection of G. oxydans 621H Zahid and Deppenmeier 2016
xynA from B. subtilis endo-1,4-β-xylanase fused N-terminally to the PelB SP PGOX0264 xylan utilization; degradation of renewable organic materials and incomplete oxidation for production of value-added products Kosciow et al. 2016
pqqA, pqqB, pqqC, pqqD, pqqE, tldD from G. oxydans WSH-003 PQQ biosynthesis proteins PpqqA, PtufB analysis of PQQ levels in G. oxydans WSH-003 and related conversion of d-sorbitol to l-sorbose Wang et al. 2016
sdhCDAB and sdhE from A. pasteurianus; sucCD from Ga. diazotrophicus succinate DH (SDH) and assembly factor SdhE for attachment of FAD to SdhA; succinyl-CoA synthetase

PGOX0264,

PsdhE

used for fragment construction for chromosomal insertion of genes to obtain G. oxydans 621H with completed TCA cycle for increased biomass yield from glucose Kiefler et al. 2017
fdhSCL from G. japonicus; membrane-bound fructose DH; extracellular sucrase SacC fused N-terminally to the PelB SP PGOX0264 production of 5-ketofructose from fructose or sucrose using genetically modified G. oxydans strains Siemen et al. 2018
sacC from Z. mobilis ZM4
fdhSCL from G. japonicus NBRC3260 membrane-bound fructose DH (Fdh) PGOX0264 production of 5-ketofructose; online monitoring of the respiration activity; pulsed and continuous fructose feed Herweg et al. 2018
phoA from E. coli; oprF from P. aeruginosa PAO1 alkaline phosphatase PhoA; first 188 aa of outer membrane porin OprF fused C-terminally to Strep-tag; OprF-PhoA fusion protein PGOX0264, PGOX0452 surface display of PhoA in G. oxydans using OprF for delivery and test of linker regions in OprF-PhoA fusion proteins Blank and Schweiger 2018
codon-optimized sldh from G. oxydans G624; lrenox from Lactobacillus reuteri sorbitol DH; NAD(P)H oxidase Padh overcoming inhibitory effect of NADPH formation during conversion of d-sorbitol to l-sorbose in G. oxydans KCTC 1091 Kim et al. 2019
fdhSCL from G. japonicus NBRC3260; inv1417 from G. japonicus LMG 1417

membrane-bound fructose DH (Fdh);

β-fructofuranosidase / invertase

PGOX0264

production of 5-ketofructose (5-KF) by G. oxydans and G. japonicus;

production of 5-KF from sucrose by invertase and Fdh with recombinant G. oxydans strains

Hoffmann et al. 2020
fdhSCL from G. japonicus NBRC3260 membrane-bound fructose DH (Fdh) PGOX0264 production of 5-ketofructose from sucrose by fructose DH and invertase with G. oxydans IK003.1 Battling et al. 2020
levS1417 from G. japonicus LMG 1417 levansucrase PGOX0264 high yield production of levan-type fructans by G. japonicus LMG 1417 Hövels et al. 2020
pBBR1MCS-4
sldAB from G. oxydans 621H membrane-bound sorbitol DH (mSLDH) PtufB, PsldAB growth performance and DHA production by engineered G. oxydans 621H Lu et al. 2012
fdhSCL from G. japonicus NBRC3260 flavoprotein-cyt. c complex fructose DH (FDH) PadhAB production of FDH in G. oxydans NBRC12528 strains; 5-keto-d-fructose formation Kawai et al. 2013
aroQ from G. oxydans NBRC3244 type II 3-dehydroquinate dehydratase Plac, ParoQ

biotransformation of quinate

to 3-dehydroshikimate by G. oxydans NBRC3244

Nishikura-Imamura et al. 2014
kgdSLC from G. oxydans NBRC3293 2-ketogluconate DH PadhAB production of 2,5-diketo-d-gluconate by G. japonicus NBRC3271 Kataoka et al. 2015
sldBA from G. frateurii NBRC101659 membrane-bound PQQ-dependent glycerol / polyol DH (GLDH) Plac sugar oxidase activities of GLDH in G. frateurii NBRC101659 strains Yakushi et al. 2018c
steP from G. frateurii NBRC101659 sugar-transporting / exporting permease none, Plac, PGOX0284, PGOX0452 SNP in steP affects trehalose efflux / sorbose uptake; NADPH/NADP+ ratio and thermotolerance of G. frateurii Matsumoto et al. 2018
quiA from G. oxydans NBRC3292 and NBRC3244 membrane-bound PQQ-dependent quinate DH (QDH) Plac, PadhAB, PquiA improving QDH activities, tested in G. oxydans NBRC12528 and G. frateurii NBRC101659 strains Yakushi et al. 2018b
sndh from Ga. liquefaciens RCTMR10 membrane-bound sorbosone DH (SNDH) PadhAB expression in G. oxydans NBRC12528 and identification of the prosthetic group of SNDH Yakushi et al. 2020

aIf no promoter or no native upstream region of the cloned gene(s) is mentioned in the publication, promoter Plac present on the pBBR1MSC-based plasmids was assumed to be responsible for target gene expression