Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Jun 30;79(3):659-668.
doi: 10.32592/ARI.2024.79.3.659. eCollection 2024 Jun.

Response Surface Methodology for Optimization of Media Components for Production of Lipase from Bacillus subtilis KUBT4

R D Nadaf  1 , P D Nadaf  2 , M M Toragall  3 , S Ct  2
Affiliations

Response Surface Methodology for Optimization of Media Components for Production of Lipase from Bacillus subtilis KUBT4

R D Nadaf et al. Arch Razi Inst. .

Abstract

Lipases are triacylglycerol hydrolases with various potential applications because of their different physical properties. Most lipase producers are extracellular in nature and are created using solid-state fermentation and submerged fermentation methods. The fungal, mycelial, and yeast lipases are produced using various solid substrates through the solid-state fermentation method. This method is cost-effective and widely used in industries to produce lipase using fungi. However, lipases from bacteria are produced using submerged fermentation. The optimization of media is a main requirement for increasing the quantitative yield by the overproduction of enzymes. The optimization of media is a main requirement for increasing the quantitative yield by overproduction of enzymes. Different parameters, such as pH, temperature, agitation speed, inoculum size, incubation time, and carbon and nitrogen sources, have been of great importance for researchers in designing economical media. The optimization by one factor at a time (OFAT) is a one-dimensional approach that is laborious and time-consuming and does not consider interactions between the factors. The limitations of OFAT method can be alleviated by employing some techniques, such as Plackett-Burman design (PBD) and response surface methodology (RSM). The PBD is a method to screen the variables that influence production and remove the non-significant factors to attain a smaller and manageable set of factors. Subsequently, the chosen significant factors are optimized by RSM that assists to study the interactions of different factors. The RSM comprises of central composite design (CCD) to fit a second-order polynomial equation. In this study, the effect of temperature, tryptone, inoculum size, and incubation time on the lipase production were analysed by PBD screening experiments. The experiments were designed using a CCD with four variables as part of RSM, utilizing the Design Expert software. This model predicted optimal activity of lipase at 58.53 U/mL when using 1.5% tryptone, a 10 mL inoculum size, and an incubation period of 48 h at 34°C. This experiment was further validated and optimal activity of lipase of 57.85 U/mL was observed. Thus, RSM model enhanced the production of lipase and can be applied for the maximum yield of lipase.

Keywords: Lipase; Plackett-Burman design; Response Surface Methodology; Bacillus subtilis KUBT4.

PubMed Disclaimer

Conflict of interest statement

Rubeen D. N., Parveen D. N., Makhadumsab M. T., and Shivasharana C. T. declare that they have no conflicts of interest.

Figures

Figure 1
Figure 1
Pareto chart showing the effect of four factors on lipase activity.
Figure 2
Figure 2
Contour plots for pectinase activity showing the interactive effects of Tryptone (%) v/s Temperature (°C).
Figure 3
Figure 3
Contour plots for pectinase activity showing the interactive effects of Inoculum size (%) v/s Temperature (°C).
Figure 4
Figure 4
Contour plots for pectinase activity showing the interactive effects of incubation time (hrs.) v/s Temperature (°C).
Figure 5
Figure 5
Contour plots for pectinase activity showing the interactive effects of Inoculum size (%) v/s Tryptone (%).
Figure 6
Figure 6
Contour plots for pectinase activity showing the interactive effects of Incubation time (hrs) v/s Tryptone (%).
Figure 7
Figure 7
Contour plots for pectinase activity showing the interactive effects of Incubation time (hrs) v/s Inoculum size (%).
Figure 8
Figure 8
Surface plots for pectinase activity showing the interactive effects of Tryptone (%) v/s Temperature (°C).
Figure 9
Figure 9
Surface plots for pectinase activity showing the interactive effects of Inoculum size (%) v/s Temperature (°C).
Figure 10
Figure 10
Surface plots for pectinase activity showing the interactive effects of Incubation time (hrs) v/s Temperature (°C).
Figure 11
Figure 11
Surface plots for pectinase activity showing the interactive effects of Inoculum size (%) v/s Tryptone (%).
Figure 12
Figure 12
Surface plots for pectinase activity showing the interactive effects of Incubation time (hrs) v/s Tryptone (%).
Figure 13
Figure 13
Surface plots for pectinase activity showing the interactive effects of Incubation time (hrs) v/s Inoculum size (%).
See this image and copyright information in PMC

References

    1. Treichel H, De Oliveira D, Mazutti MA, Di Luccio M, Oliveira JV. A review on microbial lipases production. Food Bioproc Tech. 2010;3:182–96.
    1. Colla LM, Primaz AL, Benedetti S, Loss RA, Lima MD, Reinehr CO, Bertolin TE, Costa JA. Surface response methodology for the optimization of lipase production under submerged fermentation by filamentous fungi. Braz J Microbiol. 2016;47:461–7. - PMC - PubMed
    1. Nadaf RD, Thimmappa DS. Optimization of the media components by one factor at a time methodology to enhance lipase production by Bacillus Substilis KUBT4. Int J Sci Technol Res. 2020;9(01)
    1. Palaniyappan M, Vijayagopal V, Viswanathan R, Viruthagiri T. Screening of natural substrates and optimization of operating variables on the production of pectinase by submerged fermentation using Aspergillus niger MTCC 281. Afr J Biotechnol. 2009;8(4)
    1. Montgomery DC, Keats JB, Perry LA, Thompson JR, Messina WS. Using statistically designed experiments for process development and improvement: an application in electronics manufacturing. Robot Comput Integr Manuf. 2000;16(1):55–63.

LinkOut - more resources