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Cite: NAR/gkaa742 2020



PUL General Information

Literature Information

PUL ID

PUL0454

PubMed

22944206, J Biotechnol. 2013 Aug 20;167(2):178-89. doi: 10.1016/j.jbiotec.2012.08.011. Epub 2012 Aug 31.

Characterization method

SDS-PAGE,protein fingerprinting (MALDI-TOF PMF)

Genomic accession number

CP003170.1

Nucelotide position range

4053711-4085799

Substrate

acarbose

Loci

ACPL_3663-ACPL_3684

Species

Actinoplanes sp. SE50/110/134676

Degradation or Biosynthesis

biosynthesis

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

acbZ ACPL_3663 AEV84558.1 0 - 3228 (+) CP003170.1:4053711-4056939 3.2.1.1
acbY ACPL_3664 AEV84559.1 3292 - 4096 (-) CP003170.1:4057003-4057807 -
acbX ACPL_3665 AEV84560.1 4092 - 4896 (-) CP003170.1:4057803-4058607 -
acbW ACPL_3666 AEV84561.1 4892 - 5966 (-) CP003170.1:4058603-4059677 3.6.3.31
acbV ACPL_3667 AEV84562.1 6805 - 8101 (+) CP003170.1:4060516-4061812 2.6.1.22
acbU ACPL_3668 AEV84563.1 8097 - 9576 (+) CP003170.1:4061808-4063287 -
acbS ACPL_3669 AEV84564.1 9580 - 11710 (+) CP003170.1:4063291-4065421 2.4.1.-
acbR ACPL_3670 AEV84565.1 11706 - 12798 (+) CP003170.1:4065417-4066509 2.7.7.27
acbP ACPL_3671 AEV84566.1 12794 - 13382 (+) CP003170.1:4066505-4067093 3.6.1.13
acbI ACPL_3672 AEV84567.1 13369 - 15574 (+) CP003170.1:4067080-4069285 2.4.1.-
acbJ ACPL_3673 AEV84568.1 15575 - 16427 (+) CP003170.1:4069286-4070138 3.6.1.-
acbQ ACPL_3674 AEV84569.1 16423 - 18520 (+) CP003170.1:4070134-4072231 2.4.1.25
acbK ACPL_3675 AEV84570.1 18551 - 19451 (+) CP003170.1:4072262-4073162 -
acbM ACPL_3676 AEV84571.1 19447 - 20527 (+) CP003170.1:4073158-4074238 2.7.1.-
acbL ACPL_3677 AEV84572.1 20523 - 21624 (+) CP003170.1:4074234-4075335 1.1.1.-
acbN ACPL_3678 AEV84573.1 21620 - 22376 (+) CP003170.1:4075331-4076087 1.1.1.-
acbO ACPL_3679 AEV84574.1 22372 - 23185 (+) CP003170.1:4076083-4076896 -
acbC ACPL_3680 AEV84575.1 23181 - 24378 (+) CP003170.1:4076892-4078089 4.2.3.4
acbB ACPL_3681 AEV84576.1 24438 - 25401 (-) CP003170.1:4078149-4079112 4.2.1.46
acbA ACPL_3682 AEV84577.1 25558 - 26395 (+) CP003170.1:4079269-4080106 2.7.7.24
acbE ACPL_3683 AEV84578.1 26487 - 29589 (-) CP003170.1:4080198-4083300 3.2.1.1
acbD ACPL_3684 AEV84579.1 29914 - 32089 (+) CP003170.1:4083625-4085800 2.4.1.19

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

acbZ 1 - 3228 (+) CAZyme: GH13_13|CBM41|GH13 Yes
acbY 3293 - 4096 (-) other Yes
acbX 4093 - 4896 (-) other Yes
acbW 4893 - 5966 (-) TC: gnl|TC-DB|P73329|3.A.1.141.1 Yes
acbV 6806 - 8101 (+) other Yes
acbU 8098 - 9576 (+) other Yes
acbS 9581 - 11710 (+) CAZyme: GT5|GT0 Yes
acbR 11707 - 12798 (+) other Yes
acbP 12795 - 13382 (+) STP: STP|NUDIX Yes
acbI 13370 - 15574 (+) CAZyme: GT5|GT0 Yes
acbJ 15576 - 16427 (+) other Yes
acbQ 16424 - 18520 (+) CAZyme: GH77 Yes
acbK 18552 - 19451 (+) STP: STP|PfkB,STP|PfkB Yes
acbM 19448 - 20527 (+) other Yes
acbL 20524 - 21624 (+) other Yes
acbN 21621 - 22376 (+) other Yes
acbO 22373 - 23185 (+) other Yes
acbC 23182 - 24378 (+) other Yes
acbB 24439 - 25401 (-) other Yes
acbA 25559 - 26395 (+) other Yes
acbE 26488 - 29589 (-) CAZyme: GH13_13|CBM41 Yes
acbD 29915 - 32089 (+) CAZyme: CBM20|GH13_2 Yes

PUL ID

PUL0454

PubMed

22944206, J Biotechnol. 2013 Aug 20;167(2):178-89. doi: 10.1016/j.jbiotec.2012.08.011. Epub 2012 Aug 31.

Title

The cytosolic and extracellular proteomes of Actinoplanes sp. SE50/110 led to the identification of gene products involved in acarbose metabolism.

Author

Wendler S, Hurtgen D, Kalinowski J, Klein A, Niehaus K, Schulte F, Schwientek P, Wehlmann H, Wehmeier UF, Puhler A

Abstract

The pseudotetrasaccharide acarbose is a medically relevant secondary metabolite produced by strains of the genera Actinoplanes and Streptomyces. In this study gene products involved in acarbose metabolism were identified by analyzing the cytosolic and extracellular proteome of Actinoplanes sp. SE50/110 cultures grown in a high-maltose minimal medium. The analysis by 2D protein gel electrophoresis of cytosolic proteins of Actinoplanes sp. SE50/110 resulted in 318 protein spots and 162 identified proteins. Nine of those were acarbose cluster proteins (Acb-proteins), namely AcbB, AcbD, AcbE, AcbK, AcbL, AcbN, AcbR, AcbV and AcbZ. The analysis of proteins in the extracellular space of Actinoplanes sp. SE50/110 cultures resulted in about 100 protein spots and 22 identified proteins. The identifications included the three acarbose gene cluster proteins AcbD, AcbE and AcbZ. After their identification, proteins were classified into functional groups. The dominant functional groups were the carbohydrate binding, carbohydrate cleavage and carbohydrate transport proteins. The other functional groups included protein cleavage, amino acid degradation, nucleic acid cleavage and a number of functionally uncharacterized proteins. In addition, signal peptide structures of extracellularly found proteins were analyzed. Of the 22 detected proteins 19 contained signal peptides, while 2 had N-terminal transmembrane helices explaining their localization. The only protein having neither of them was enolase. Under the conditions applied, the secretome of Actinoplanes sp. SE50/110 was dominated by seven proteins involved in carbohydrate metabolism (PulA, AcbE, AcbD, MalE, AglE, CbpA and Cgt). Of special interest were the identified extracellular pullulanase PulA and the two solute-binding proteins MalE and AglE. The identifications suggest that Actinoplanes sp. SE50/110 has two maltose/maltodextrin import systems. We postulate the identified MalEFG transport system of Actinoplanes sp. SE50/100 as the missing acarbose-metabolite importer and present a model of acarbose metabolism that is extended by the newly identified gene products.