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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.

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Related in: MedlinePlus

Physiological analyses of the S. lividansΔH transformants complemented with a different plasmid type. The ΔH strains with the control plasmid pWHM3 (A, E and I), with pHY11 (B, F and J), pHY12 (C, G and K) or with pHY13 (D, H and L) were grown for 19 h, and subsequently shaken for 20 h (A–D), 40 h (E–H) or 120 h (I–L). The samples were analysed by phase‐contrast microscopy. The magnification of the pictures (A)–(D) (bar in D), (E)–(H) (bar in H) and (I)–(L) (bar in L) is indicated.
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f8: Physiological analyses of the S. lividansΔH transformants complemented with a different plasmid type. The ΔH strains with the control plasmid pWHM3 (A, E and I), with pHY11 (B, F and J), pHY12 (C, G and K) or with pHY13 (D, H and L) were grown for 19 h, and subsequently shaken for 20 h (A–D), 40 h (E–H) or 120 h (I–L). The samples were analysed by phase‐contrast microscopy. The magnification of the pictures (A)–(D) (bar in D), (E)–(H) (bar in H) and (I)–(L) (bar in L) is indicated.

Mentions: As turbidity measurements are not conclusive to investigate cultures of Streptomyces mycelia, pre‐cultivated strains were transferred to optimized shaking conditions (145 r.p.m.), and the flasks were inspected microscopically. After 20 h, the substrate hyphae of ΔHpHY11, ΔHpHY12 and ΔHpHY13 (Fig. 8B–D) formed more fused pellets compared with those the control ΔHpWHM3 strain (Fig. 8A). Pellets of ΔHpHY12 and ΔHpHY13 were more roundish. The pigmentation of all strains appeared similar. After further 40 h of cultivation, the pellets of ΔHpHY13 (Fig. 8H) were the most irregular as to shape and size, and they had more fused packages than those of ΔHpHY11 (Fig. 8F). Pellets of ΔHpHY12 (Fig. 8G) had the smallest diameter, and they comprised roundish and elongated ones. In addition, they had fewer fusion sites than those of ΔHpHY11. The aggregates of the control strain ΔHpWHM3 (Fig. 8E) had larger diameters than those of ΔHpHY11 and ΔHpHY12 and lacked close contact sites. The degree of pigmentation appeared most pronounced for ΔHpHY11 and ΔHpHY13. Following prolonged cultivation (120 h) the pellets of ΔHpHY12 (Fig. 8K) and ΔHpWHM3 (Fig. 8I) were transparent, and their diameter diminished compared with 40 h. In addition, a pronounced cloudy‐like background was present; this was due to masses of individual hyphae and small mycelia (Fig. 8I and K). The pellet clumps of ΔHpHY13 (Fig. 8L) were more compact, and patches within them were more coloured compared with those of ΔHpHY11 (Fig. 8J). These features correlated with fewer individual hyphae and small mycelia. Taken together, the pellets of ΔHpHY13 were even more stable than those ΔHpHY11.


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)

Physiological analyses of the S. lividansΔH transformants complemented with a different plasmid type. The ΔH strains with the control plasmid pWHM3 (A, E and I), with pHY11 (B, F and J), pHY12 (C, G and K) or with pHY13 (D, H and L) were grown for 19 h, and subsequently shaken for 20 h (A–D), 40 h (E–H) or 120 h (I–L). The samples were analysed by phase‐contrast microscopy. The magnification of the pictures (A)–(D) (bar in D), (E)–(H) (bar in H) and (I)–(L) (bar in L) is indicated.
© Copyright Policy
Related In: Results  -  Collection

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

f8: Physiological analyses of the S. lividansΔH transformants complemented with a different plasmid type. The ΔH strains with the control plasmid pWHM3 (A, E and I), with pHY11 (B, F and J), pHY12 (C, G and K) or with pHY13 (D, H and L) were grown for 19 h, and subsequently shaken for 20 h (A–D), 40 h (E–H) or 120 h (I–L). The samples were analysed by phase‐contrast microscopy. The magnification of the pictures (A)–(D) (bar in D), (E)–(H) (bar in H) and (I)–(L) (bar in L) is indicated.
Mentions: As turbidity measurements are not conclusive to investigate cultures of Streptomyces mycelia, pre‐cultivated strains were transferred to optimized shaking conditions (145 r.p.m.), and the flasks were inspected microscopically. After 20 h, the substrate hyphae of ΔHpHY11, ΔHpHY12 and ΔHpHY13 (Fig. 8B–D) formed more fused pellets compared with those the control ΔHpWHM3 strain (Fig. 8A). Pellets of ΔHpHY12 and ΔHpHY13 were more roundish. The pigmentation of all strains appeared similar. After further 40 h of cultivation, the pellets of ΔHpHY13 (Fig. 8H) were the most irregular as to shape and size, and they had more fused packages than those of ΔHpHY11 (Fig. 8F). Pellets of ΔHpHY12 (Fig. 8G) had the smallest diameter, and they comprised roundish and elongated ones. In addition, they had fewer fusion sites than those of ΔHpHY11. The aggregates of the control strain ΔHpWHM3 (Fig. 8E) had larger diameters than those of ΔHpHY11 and ΔHpHY12 and lacked close contact sites. The degree of pigmentation appeared most pronounced for ΔHpHY11 and ΔHpHY13. Following prolonged cultivation (120 h) the pellets of ΔHpHY12 (Fig. 8K) and ΔHpWHM3 (Fig. 8I) were transparent, and their diameter diminished compared with 40 h. In addition, a pronounced cloudy‐like background was present; this was due to masses of individual hyphae and small mycelia (Fig. 8I and K). The pellet clumps of ΔHpHY13 (Fig. 8L) were more compact, and patches within them were more coloured compared with those of ΔHpHY11 (Fig. 8J). These features correlated with fewer individual hyphae and small mycelia. Taken together, the pellets of ΔHpHY13 were even more stable than those ΔHpHY11.

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