Basic Study
Copyright ©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Biol Chem. Aug 26, 2015; 6(3): 265-271
Published online Aug 26, 2015. doi: 10.4331/wjbc.v6.i3.265
Effect of agitation speed on the morphology of Aspergillus niger HFD5A-1 hyphae and its pectinase production in submerged fermentation
Darah Ibrahim, Haritharan Weloosamy, Sheh-Hong Lim
Darah Ibrahim, Haritharan Weloosamy, Sheh-Hong Lim, Industrial Biotechnology Research Laboratory, School of Biological Sciences, Universiti Sains Malaysia, 11800 Penang, Malaysia
Author contributions: Ibrahim D designed and coordinated the research; Weloosamy H performed the majority of experiments; Lim SH analyzed the data; Ibrahim D, Weloosamy H and Lim SH wrote the paper.
Supported by Universiti Sains Malaysia Scientific Research Fund.
Institutional review board statement: Our experiment was not related to the stated fields. Thus, no document can be provided.
Conflict-of-interest statement: No conflict of interest to declare.
Data sharing statement: We do not have data sharing statement to be attached.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Sheh-Hong Lim, PhD, Researcher, Industrial Biotechnology Research Laboratory, School of Biological Sciences, Universiti Sains Malaysia, 11800 Penang, Malaysia. limshehhong77@gmail.com
Telephone: +60-12-4629487 Fax: +60-4-6565125
Received: March 5, 2015
Peer-review started: March 7, 2015
First decision: April 27, 2015
Revised: June 7, 2015
Accepted: June 30, 2015
Article in press: July 2, 2015
Published online: August 26, 2015
Abstract

AIM: To investigate the impact of agitation speed on pectinase production and morphological changing of Aspergillus niger (A. niger) HFD5A-1 in submerged fermentation.

METHODS: A. niger HFM5A-1 was isolated from a rotted pomelo. The inoculum preparation was performed by adding 5.0 mL of sterile distilled water containing 0.1% Tween 80 to a sporulated culture. Cultivation was carried out with inoculated 1 × 107 spores/mL suspension and incubated at 30 °C with different agitation speed for 6 d. The samples were withdrawn after 6 d cultivation time and were assayed for pectinase activity and fungal growth determination. The culture broth was filtered through filter paper (Whatman No. 1, London) to separate the fungal mycelium. The cell-free culture filtrate containing the crude enzyme was then assayed for pectinase activity. The biomass was dried at 80 °C until constant weight. The fungal cell dry weight was then expressed as g/L. The 6 d old fungal mycelia were harvested from various agitation speed, 0, 50, 100, 150, 200 and 250 rpm. The morphological changing of samples was then viewed under the light microscope and scanning electron microscope.

RESULTS: In the present study, agitation speed was found to influence pectinase production in a batch cultivation system. However, higher agitation speeds than the optimal speed (150 rpm) reduced pectinase production which due to shear forces and also collision among the suspended fungal cells in the cultivation medium. Enzyme activity increased with the increasing of agitation speed up to 150 rpm, where it achieved its maximal pectinase activity of 1.559 U/mL. There were significant different (Duncan, P < 0.05) of the pectinase production with the agitation speed at static, 50, 100, 200 and 250 rpm. At the static condition, a well growth mycelial mat was observed on the surface of the cultivation medium and sporulation occurred all over the fungal mycelial mat. However with the increased in agitation speed, the mycelial mat turned slowly to become a single circular pellet. Thus, it was found that agitation speed affected the morphological characteristics of the fungal hyphae/mycelia of A. niger HFD5A-1 by altering their external as well as internal cell structures.

CONCLUSION: Exposure to higher shear stress with an increasing agitation speed could result in lower biomass yields as well as pectinase production by A. niger HFD5A-1.

Keywords: Aspergillus niger, Agitation, Submerged culture, Pectinase, Microbial growth, Dissolved oxygen

Core tip: We report the influence of agitation speed on pectinase production on our newly isolate Aspergillus niger (A. niger) HFM5A-1 in submerged fermentation. The agitation speed was found to influence pectinase production in a batch cultivation system. However, higher agitation speeds than the optimal speed (150 rpm) reduced pectinase production which due to shear forces and also collision among the suspended fungal cells in the cultivation medium. It was found that agitation speed affected the morphological characteristics of the fungal hyphae/mycelia of A. niger HFD5A-1 by altering their external as well as internal cell structures.