Abstract
Mycelial morphogenesis and the production of fungal secretory proteins are still largely unknown. A mutant strain of Aspergillus carbonarius UV-10046 produced abundant polygalacturonase (PG) along with partially saturated canthaxanthin (PSC) at low pH conditions. In the present study, the relationship between PG secretion and PSC biosynthesis was studied using carotenogenic inhibitors and SDS-PAGE electrophoresis. Also the correlation between morphogenesis and PG secretion was investigated by analysing through microscopic studies. From the results, it was observed that secretion of PG was positively influenced by the PSC biosynthesis. The results also showed that the mutant with hairy mycelial structure resulted in higher PG activity when compared to the wild type that lacks hyper branching. From the results, it was confirmed that a mutation might have occurred in the isoprenoid pathway that has helped mutant for survival at acidic conditions. Further, an alteration in the morphogenesis and hyper branching development caused over secretion of PG enzyme in the mutant.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aguiar TQ, Ribeiro O, Arvas M, Wiebe MG, Penttilä M, Domingues L (2014) Investigation of protein secretion and secretion stress in Ashbya gossypii. BMC Genomics 15:1137. https://doi.org/10.1186/1471-2164-15-1137
Ahamed A, Vermette P (2009) Effect of culture medium composition on Trichoderma reesei’s morphology and cellulase production. Bioresour Technol 100:5979–5987. https://doi.org/10.1016/j.biortech.2009.02.070
Ahamed A, Vermette P (2010) Effect of mechanical agitation on the production of cellulases by Trichoderma reesei RUT-C30 in a draft-tube airlift bioreactor. Biochem Eng 49:379–387. https://doi.org/10.1016/j.bej.2010.01.014
Amanullah A, Christensen L, Hansen K, Nienow A, Thomas C (2002) Dependence of morphology on agitation intensity in fed-batch cultures of Aspergillus oryzae and its implications for recombinant protein production. Biotechnol Bioeng 77:815–826. https://doi.org/10.1002/bit.10181
An G-H, Jang B-G, Cho M-H (2001) Cultivation of the carotenoid-hyperproducing mutant 2A2N of the red yeast Xanthophyllomyces dendrorhous (Phaffia rhodozyma) with molasses. J Biosci Bioeng 92:121–125. https://doi.org/10.1263/jbb.92.121
Braun S, Vecht-Lifshitz SE (1991) Mycelial morphology and metabolite production. Trends Biotechnol 9:63–68. https://doi.org/10.1016/0167-7799(91)90020-I
Dubey A, Suresh C, Kavitha R, Karanth N, Umesh-Kumar S (2000) Evidence that the glucoamylases and α-amylase secreted by Aspergillus niger are proteolytically processed products of a precursor enzyme. FEBS Lett 471:251–255. https://doi.org/10.1016/S0014-5793(00)01410-1
Gerlach S, Siedenberg D, Gerlach D, Schügerl K, Giuseppin M, Hunik J (1998) Influence of reactor systems on the morphology of Aspergillus awamori. Application of neural network and cluster analysis for characterization of fungal morphology. Process Biochem 33:601–615. https://doi.org/10.1016/S0032-9592(98)00016-8
Gordon C et al (2000) A glucoamylase: GFP gene fusion to study protein secretion by individual hyphae of Aspergillusniger. J Microbiol Methods 42:39–48. https://doi.org/10.1016/S0167-7012(00)00170-6
Gulati R, Isar J, Kumar V, Prasad AK, Parmar VS, Saxena RK (2005) Production of a novel alkaline lipase by Fusarium globulosum using neem oil, and its applications. Pure Appl Chem 77:251–262. https://doi.org/10.1351/pac200577010251
Haack MB, Olsson L, Hansen K, Lantz AE (2006) Change in hyphal morphology of Aspergillus oryzae during fed-batch cultivation. Appl Microbiol Biotechnol 70:482–487. https://doi.org/10.1007/BF00253786
He R, Li C, Ma L, Zhang D, Chen S (2016) Effect of highly branched hyphal morphology on the enhanced production of cellulase in Trichoderma reesei DES-15. 3. Biotech 6:214. https://doi.org/10.1007/s13205-016-0516-5
Horowitz W (1980) Official methods of analysis of the association of analytical chemists. J Assoc Offic Anal Chem, Washington
Hwang S-W, Choi HI, Sim SJ (2019) Acidic cultivation of Haematococcus pluvialis for improved astaxanthin production in the presence of a lethal fungus. Bioresour Technol 278:138–144. https://doi.org/10.1016/j.biortech.2019.01.080
Ikram-ul-Haq S, Muhammad M, Zafar S, Tehmina S (2006) Triggering of beta-glucosidase production in Trichoderma viride with nutritional and environmental control. J Appl Sci Res 2:884–889
Kumar A, Mathimaran A, Shrikanta A, Govindaswamy V (2018) Role of partially saturated canthaxanthin and ergosterol in the survival of Aspergillus carbonarius mutant at extreme acidic condition. Microbiology 87:183–190. https://doi.org/10.1134/S0026261718020066
Kumar A, Srikanta AH, Chinnaswamy A, Lai OM, Govindaswamy V (2016) Encapsulation of partially saturated canthaxanthin-A functional ingredient from Aspergillus carbonarius mutant. Curr Top Nutraceutical Res 14:251–258
Kumaresan N, Sanjay KR, Venkatesh KS, Kadeppagari R-K, Vijayalakshmi G, Umesh-Kumar S (2008) Partially saturated canthaxanthin purified from Aspergillus carbonarius induces apoptosis in prostrate cancer cell line. Appl Microbiol Biotechnol 80:467–473. https://doi.org/10.1007/s00253-008-1538-7
Laemmli U (1970) Most commonly used discontinuous buffer system for SDS electrophoresis. Nature 227:680–686
Luo H, Niu Y, Duan C, Su H, Yan G (2013) A pH control strategy for increased β-carotene production during batch fermentation by recombinant industrial wine yeast. Process Biochem 48:195–200. https://doi.org/10.1016/j.procbio.2013.01.005
Miles EA, Trinci A (1983) Effect of pH and temperature on morphology of batch and chemostat cultures of Penicillium chrysogenum. Trans Brit 81:193–200. https://doi.org/10.1016/S0007-1536(83)80069-2
Morrissey JH (1981) Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem 117:307–310. https://doi.org/10.1016/0003-2697(81)90783-1
Müller C, McIntyre M, Hansen K, Nielsen J (2002) Metabolic engineering of the morphology of Aspergillus oryzae by altering chitin synthesis. Appl Environ Microbiol 68:1827–1836. https://doi.org/10.1128/AEM.68.4.1827-1836.2002
Müller C, Spohr AB, Nielsen J (2000) Role of substrate concentration in mitosis and hyphal extension of Aspergillus. Biotechnol Bioeng 67:390–397. https://doi.org/10.1002/(sici)1097-0290(20000220)67:4%3c390:aid-bit2%3e3.0.co;2-o
Norouzian D (2008) Effect of different factors on fermentative production of enzymes by fungi. Dyn Biochem Process Biotechnol Mol Biol 2:14–18
Papagianni M (2004) Fungal morphology and metabolite production in submerged mycelial processes. Biotechnol Adv 22:189–259. https://doi.org/10.1016/j.biotechadv.2003.09.005
Rahardjo YSP (2005) Fungal mats in solid-state fermentation. Wageningen University, Wageningen
Rao P, Suresh C, Rao DN, Kumar SU, Divakar S (1999) Digestion of residual β-cyclodextrin in treated egg using glucoamylase from a mutant strain of Aspergillus niger (CFTRI 1105). Food Chem 65:297–301. https://doi.org/10.1016/S0308-8146(98)00189-7
Ravi-Kumar K, Umesh-Kumar S (2005) 06 Bioprocessing of Starch Using Enzyme Technology. Food Biotechnology. CRC Press, Boca Raton, pp 736–749. https://doi.org/10.1201/9781420027976.ch2.06
Ravi-Kumar K, Venkatesh K, Umesh-Kumar S (2004) Evidence that cleavage of the precursor enzyme by autocatalysis caused secretion of multiple amylases by Aspergillus niger. FEBS Lett 557:239–242. https://doi.org/10.1016/S0014-5793(03)01510-2
Ravi-Kumar K, Venkatesh K, Umesh-Kumar S (2007) The 53-kDa proteolytic product of precursor starch-hydrolyzing enzyme of Aspergillus niger has Taka-amylase-like activity. Appl Microbiol Biotechnol 74:1011–1015. https://doi.org/10.1007/s00253-006-0742-6
Shubakov AA, Elkina EA (2002) Production of polygalacturonases by filamentous fungi Aspergillus niger ACMF-1119 and Penicillium dierckxii ACIMF-152. Chemistry and Computational Simulation 2:65–68
Smith J, Lilly M, Fox R (1990) The effect of agitation on the morphology and penicillin production of Penicillium chrysogenum. Biotechnol Bioeng 35:1011–1023. https://doi.org/10.1002/bit.260351009
Somogyi M (1952) Notes on sugar determination. J Biol Chem 195:19–23
Spector T (1978) Refinement of the Coomassie blue method of protein quantitation: a simple and linear spectrophotometric assay for ≤ 0.5 to 50 μg of protein. Anal Biochem 86:142–146. https://doi.org/10.1016/0003-2697(78)90327-5
Suresh C, Dubey A, Kavitha R, Umesh-Kumar S (1998) An immunoaffinity procedure for determination of enzyme activity at elevated temperatures. Anal Biochem 264:286–288. https://doi.org/10.1006/abio.1998.2867
Suresh C, Dubey A, Srikanta S, Kumar SU, Karanth N (1999a) Characterisation of a starch-hydrolysing enzyme of Aspergillus niger. Appl Microbiol Biotechnol 51:673–675. https://doi.org/10.1002/(SICI)1522-2683(19990301)20:3%3C483:AID-ELPS483%3E3.0.CO;2-B
Suresh C, Dubey AK, Kini R, Umesh-Kumar S, Karanth NG (1999b) Separation and direct detection of raw and gelatinized starch hydrolyzing activities of glucoamylase on isoelectric focusing gels. Electrophoresis 20:483–485. https://doi.org/10.1007/s002530051450
Te Biesebeke R et al (2005) Branching mutants of Aspergillus oryzae with improved amylase and protease production on solid substrates. Appl Microbiol Biotechnol 69:44. https://doi.org/10.1007/s00253-005-1968-4
Unluturk S, Oncu S, Tari C, Gogus N (2006) Various factors affecting the pellet morphology, broth reology and pectinase enzyme production in submerged fermentation of Aspergillus sojae. In: 13th IUFoST 2006, pp 271–271 doi: 10.1051/IUFoST:20060271
Vecht-Lifshitz S, Magdassi S, Braun S (1990) Pellet formation and cellular aggregation in Streptomyces tendae. Biotechnol Bioeng 35:890–896. https://doi.org/10.1002/bit.260350906
Wardell JN, Bushell ME (1999) Kinetics and manipulation of hyphal breakage and its effect on antibiotic production. Enzyme Microb Technol 25:404–410. https://doi.org/10.1016/S0141-0229(99)00064-2
Wessels JG (1993) Tansley review no. 45 wall growth, protein excretion and morphogenesis in fungi. New Phytol 123:397–413. https://doi.org/10.1111/j.1469-8137.1993.tb03751.x
Whitaker A, Long P (1973) Fungal pelleting. Process Biochem 8:27–31
Wongwicharn A, Harvey LM, McNeil B (1999a) Secretion of heterologous and native proteins, growth and morphology in batch cultures of Aspergillus niger B1-D at varying agitation rates. J Chem Technol Biotechnol Int Res Process, Environ Clean Technol 74:821–828. https://doi.org/10.1002/(SICI)1097-4660(199908)74:8%3c821:AID-JCTB114%3e3.0.CO;2-%23
Wongwicharn A, McNeil B, Harvey LM (1999b) Effect of oxygen enrichment on morphology, growth, and heterologous protein production in chemostat cultures of Aspergillus niger B1-D. Biotechnol Bioeng 65:416–424. https://doi.org/10.1002/(SICI)1097-0290(19991120)65:4%3c416:AID-BIT6%3e3.0.CO;2-Z
Wösten HA, Moukha SM, Sietsma JH, Wessels JG (1991) Localization of growth and secretion of proteins in Aspergillus niger. Microbiology 137:2017–2023. https://doi.org/10.1099/00221287-137-8-2017
Xiang X, Morris NR (1999) Hyphal tip growth and nuclear migration. Curr Opin Microbiol 2:636–640. https://doi.org/10.1016/S1369-5274(99)00034-X
Acknowledgements
This work was supported by the grants from the Department of Science and Technology (DST), Science and Engineering Research board (SERB), India, under Young Scientist Scheme (Project No. YSS/2015/000298).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no conflict of interest.
Additional information
Communicated by Erko Stackebrandt.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Mathimaran, A., Kumar, A. Changes in morphogenesis and carotenogenesis to influence polygalacturonase secretion in Aspergillus carbonarius mutant. Arch Microbiol 202, 1285–1293 (2020). https://doi.org/10.1007/s00203-020-01838-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00203-020-01838-7