A molecular key for building hyphae aggregates: the role of the newly identified Streptomyces protein HyaS.
Streptomycetes produce many metabolites with medical and biotechnological applications. During fermentations, their hyphae build aggregates, a process in which the newly identified protein HyaS plays an important role. The corresponding hyaS gene is present within all investigated Streptomyces species. Reporter fusions indicate that transcription of hyaS occurs within substrate hyphae of the Streptomyces lividans wild type (WT). The HyaS protein is dominantly associated with the substrate hyphae. The WT strain forms cylindrically shaped clumps of densely packed substrate hyphae, often fusing to higher aggregates (pellets), which remain stably associated during shaking. Investigations by electron microscopy suggest that HyaS induces tight fusion-like contacts among substrate hyphae. In contrast, the pellets of the designed hyaS disruption mutant ΔH are irregular in shape, contain frequently outgrowing bunches of hyphae, and fuse less frequently. ΔH complemented with a plasmid carrying hyaS resembles the WT phenotype. Biochemical studies indicate that the C-terminal region of HyaS has amine oxidase activity. Investigations of ΔH transformants, each carrying a specifically mutated gene, lead to the conclusion that the in situ oxidase activity correlates with the pellet-inducing role of HyaS, and depends on the presence of certain histidine residues. Furthermore, the level of undecylprodigiosin, a red pigment with antibiotic activity, is influenced by the engineered hyaS subtype within a strain. These data present the first molecular basis for future manipulation of pellets, and concomitant production of secondary metabolites during biotechnological processes.
University of Osnabrück, FB Biology/Chemistry, Applied Genetics of Microorganisms, 49069 Osnabrück, Germany.
Ilona
I
Koebsch
Jens
J
Overbeck
Sophie
S
Piepmeyer
Holger
H
Meschke
Hildgund
H
Schrempf
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D025301:Hyphae / Q000737:chemistry / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D008969:Molecular Sequence Data; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D048170:Streptomyces lividans / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development
D001426:Bacterial Proteins; D000587:Oxidoreductases Acting on CH-NH Group Donors; C019668:amine oxidase, mixed function
Biosci Biotechnol Biochem. 2007 Feb;71(2):283-99
17284841
Arch Microbiol. 2008 Aug;190(2):119-27
18438642
J Bacteriol. 1989 Nov;171(11):5872-81
2681144
J Basic Microbiol. 1992;32(3):193-200
1512710
Anal Biochem. 1984 Jan;136(1):175-9
6424501
J Bacteriol. 1987 Oct;169(10):4804-10
2820944
Mol Microbiol. 1992 Dec;6(23):3611-21
1282194
Nucleic Acids Res. 1997 Sep 1;25(17):3389-402
9254694
Curr Opin Microbiol. 2003 Dec;6(6):564-71
14662351
Nature. 1970 Aug 15;227(5259):680-5
5432063
Appl Microbiol Biotechnol. 2000 Jul;54(1):33-6
10952002
Bioprocess Biosyst Eng. 2005 Apr;27(2):99-104
15592878
Mol Microbiol. 2006 Feb;59(3):731-42
16420347
Annu Rev Microbiol. 1981;35:365-403
6170248
Nat Biotechnol. 2003 May;21(5):526-31
12692562
Eur J Biochem. 1997 Jun 1;246(2):557-64
9208950
Arch Microbiol. 2005 Feb;183(2):95-106
15599705
BMC Microbiol. 2006 Jun 27;6:59
16803626
Gene. 1997 May 6;190(2):315-7
9197550
Appl Microbiol Biotechnol. 2002 Dec;60(4):377-80
12466876
Microbiology (Reading). 2004 Jan;150(Pt 1):21-31
14702394
Protein Eng. 1997 Jan;10(1):1-6
9051728
EMBO J. 2001 Aug 1;20(15):3893-901
11483492
J Bacteriol. 2008 Jun;190(11):4050-60
18375553
Biotechnol Bioeng. 2005 Dec 5;92(5):614-23
16136592
Gene. 1982 Oct;19(3):259-68
6295879
Mol Microbiol. 2006 Apr;60(1):5-15
16556216
Nucleic Acids Res. 1992 Mar 11;20(5):961-74
1549509
Biotechnol Prog. 2007 Jul-Aug;23(4):836-45
17585778
FEMS Microbiol Lett. 2006 Mar;256(1):1-15
16487313
J Histochem Cytochem. 1981 May;29(5):663-71
6166664
Nature. 2002 May 9;417(6885):141-7
12000953
Plant Physiol. 2002 Mar;128(3):865-75
11891243
Biosci Biotechnol Biochem. 2007 Jun;71(6):1373-86
17587668
J Microbiol Biotechnol. 2007 Dec;17(12):2066-70
18167457
J Microsc. 2008 Jan;229(Pt 1):174-82
18173655
Appl Microbiol Biotechnol. 2006 Nov;73(1):1-14
17013601
Comput Appl Biosci. 1992 Apr;8(2):189-91
1591615
Plasmid. 1995 Mar;33(2):113-26
7597106
Nat Biotechnol. 2006 Dec;24(12):1533-40
17160059
J Biol Chem. 1972 Apr 10;247(7):2212-6
4552692
J Biotechnol. 2002 May 9;95(2):133-44
11911923
Blood. 1982 Nov;60(5):1195-202
7126871