Limits...
A molecular key for building hyphae aggregates: the role of the newly identified Streptomyces protein HyaS.

Koebsch I, Overbeck J, Piepmeyer S, Meschke H, Schrempf H - Microb Biotechnol (2009)

Bottom Line: The HyaS protein is dominantly associated with the substrate hyphae.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.These data present the first molecular basis for future manipulation of pellets, and concomitant production of secondary metabolites during biotechnological processes.

View Article: PubMed Central - PubMed

Affiliation: University of Osnabrück, FB Biology/Chemistry, Applied Genetics of Microorganisms, 49069 Osnabrück, Germany.

Show MeSH

Related in: MedlinePlus

Characteristics of pellets from shaking cultures of S. lividans WT and ΔH. The WT (A and B, and E and F) strain or the ΔH mutant (C and D, and G and H) were grown in complete medium after shaking for 7 h (A and C) or 17 h (B, D and E; G, F and H), and inspected microscopically under visual light by phase contrast (A–G). After treatment with primary anti‐HyaS antibodies followed by Alexa Fluor‐coupled secondary anti‐rat antibodies, samples (E and G) were analysed under UV light with a Cy5 filter (F and H). The magnification of the pictures (A)–(D) (see bar in D) differs from that of the pictures (E)–(H) (see bar in H).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3815755&req=5

f4: Characteristics of pellets from shaking cultures of S. lividans WT and ΔH. The WT (A and B, and E and F) strain or the ΔH mutant (C and D, and G and H) were grown in complete medium after shaking for 7 h (A and C) or 17 h (B, D and E; G, F and H), and inspected microscopically under visual light by phase contrast (A–G). After treatment with primary anti‐HyaS antibodies followed by Alexa Fluor‐coupled secondary anti‐rat antibodies, samples (E and G) were analysed under UV light with a Cy5 filter (F and H). The magnification of the pictures (A)–(D) (see bar in D) differs from that of the pictures (E)–(H) (see bar in H).

Mentions: To investigate the role of the hyaS gene within the WT chromosome (Fig. 3, line D), we constructed several independent S. lividans mutants (named ΔH, Fig. 3, line E, and Table 1), which carried a hyaS gene disrupted by the hygromycin‐resistance cassette (Ωhyg) as outlined under Experimental procedures. On solid medium, ΔH colonies grew like the WT strain, and shared the differentiation pattern leading to spores (not shown). As viewed by transmission‐electron microscopy, the shape of the sectioned mutant spores corresponded to those of the WT strain (following Figs 4 and 5, the spores are presented in Fig. 6A and B).


A molecular key for building hyphae aggregates: the role of the newly identified Streptomyces protein HyaS.

Koebsch I, Overbeck J, Piepmeyer S, Meschke H, Schrempf H - Microb Biotechnol (2009)

Characteristics of pellets from shaking cultures of S. lividans WT and ΔH. The WT (A and B, and E and F) strain or the ΔH mutant (C and D, and G and H) were grown in complete medium after shaking for 7 h (A and C) or 17 h (B, D and E; G, F and H), and inspected microscopically under visual light by phase contrast (A–G). After treatment with primary anti‐HyaS antibodies followed by Alexa Fluor‐coupled secondary anti‐rat antibodies, samples (E and G) were analysed under UV light with a Cy5 filter (F and H). The magnification of the pictures (A)–(D) (see bar in D) differs from that of the pictures (E)–(H) (see bar in H).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3815755&req=5

f4: Characteristics of pellets from shaking cultures of S. lividans WT and ΔH. The WT (A and B, and E and F) strain or the ΔH mutant (C and D, and G and H) were grown in complete medium after shaking for 7 h (A and C) or 17 h (B, D and E; G, F and H), and inspected microscopically under visual light by phase contrast (A–G). After treatment with primary anti‐HyaS antibodies followed by Alexa Fluor‐coupled secondary anti‐rat antibodies, samples (E and G) were analysed under UV light with a Cy5 filter (F and H). The magnification of the pictures (A)–(D) (see bar in D) differs from that of the pictures (E)–(H) (see bar in H).
Mentions: To investigate the role of the hyaS gene within the WT chromosome (Fig. 3, line D), we constructed several independent S. lividans mutants (named ΔH, Fig. 3, line E, and Table 1), which carried a hyaS gene disrupted by the hygromycin‐resistance cassette (Ωhyg) as outlined under Experimental procedures. On solid medium, ΔH colonies grew like the WT strain, and shared the differentiation pattern leading to spores (not shown). As viewed by transmission‐electron microscopy, the shape of the sectioned mutant spores corresponded to those of the WT strain (following Figs 4 and 5, the spores are presented in Fig. 6A and B).

Bottom Line: The HyaS protein is dominantly associated with the substrate hyphae.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.These data present the first molecular basis for future manipulation of pellets, and concomitant production of secondary metabolites during biotechnological processes.

View Article: PubMed Central - PubMed

Affiliation: University of Osnabrück, FB Biology/Chemistry, Applied Genetics of Microorganisms, 49069 Osnabrück, Germany.

Show MeSH
Related in: MedlinePlus