|Astri Nur Istyami||Department of Bioenergy Engineering and Chemurgy, Institut Teknologi Bandung, Jl. Raya Jatinangor KM 20.75, Kabupaten Sumedang 45363, Indonesia|
|Ronny Purwadi||Department of Food Engineering, Institut Teknologi Bandung, Jl. Raya Jatinangor KM 20.75, Kabupaten Sumedang 45363, Indonesia|
|Made Tri Ari Penia Kresnowati||Department of Food Engineering, Institut Teknologi Bandung, Jl. Raya Jatinangor KM 20.75, Kabupaten Sumedang 45363, Indonesia|
|Tirto Prakoso||Department of Bioenergy Engineering and Chemurgy, Institut Teknologi Bandung, Jl. Raya Jatinangor KM 20.75, Kabupaten Sumedang 45363, Indonesia|
|Tatang Hernas Soerawidjaja||Department of Bioenergy Engineering and Chemurgy, Institut Teknologi Bandung, Jl. Raya Jatinangor KM 20.75, Kabupaten Sumedang 45363, Indonesia|
Free fatty acid, which is an important intermediate product in the oleochemical industry, can be produced by hydrolysis of oil using lipase enzymes. This process is more economical and less energy consuming than the conventional process, i.e. noncatalytic thermal hydrolysis. While lipase from microorganisms requires a complex separation step, that from plants involves lower cost and easier handling. Nevertheless, no report has been published on the immobilization of plant latex-based lipase, while immobilization to increase the economic feasibility of microbial lipases has been widely reported. The aim of this study is to compare the performance of free and immobilized frangipani latex lipase in palm oil lipolysis. Immobilization was conducted by physical adsorption using hydrophobic supports and matrix encapsulation. The adsorption of frangipani latex lipase onto polypropylene and polyethylene beads was found to be ineffective, although the presence of the beads did slightly increase the degree of lipolysis. On the other hand, encapsulation with a calcium alginate matrix was effective in immobilizing particulate latex, although the calcium alginate beads were susceptible to breaking, causing contamination of the lipolysis product. To develop lipolysis technology utilizing frangipani latex lipase, free form lipase is more suitable in small-scale, stirred-tank lipolysis, while lipolysis with immobilized lipase from frangipani latex requires further modification, such as use of a packed bed reactor, circulated flow, or matrix modification.
Fatty acids; Frangipani; Immobilized lipase; Latex lipase; Lipolysis
As the world is showing great interest in sustainable industry, demand for oleochemical products has increased in the last decades. These products are slowly replacing petrochemical ones, including surfactants, plastics, lubricants, and even fuels. One of the most important reactions involved in the oleochemical industry is the conversion of triglyceride into fatty acids, which is encountered in most plant oil processing into derivative products. With the potential for increasing demand in the future, it is necessary to ensure that fatty acid production technology is energy-efficient, cost-efficient, and effective.
Noncatalytic thermal hydrolysis of triglyceride is the current technology employed for fatty acid production. It is a robust (260oC, 50 bar) and high-energy-consuming process (Barnebey & Brown, 1948). The high temperature of the hydrolysis triggers unwanted reactions (Mounguengui et al., 2013), so a process involving milder conditions is preferable to avoid these drawbacks.
Lipase (EC 22.214.171.124) is an enzyme which catalyzes triglyceride hydrolysis (or lipolysis, when lipase is used). Lipases from bacteria and fungi have been studied for a long time, and some have been commercially produced. Although they are readily available in large quantities, their application in industrial lipolysis is limited by their high production cost (Seth et al., 2014). Other natural sources of lipase have emerged as alternatives; for example, plant seeds (Barros et al., 2010) and plant latex (Mazou et al., 2016). One remarkably active lipase source is frangipani (Plumeria rubra) latex particulate (Cambon et al., 2006). With the abundance of frangipani trees in warm regions, it is potentially feasible to develop small-scale production plants of fatty acids in rural areas.
Immobilization techniques has been utilized to improve the economic feasibility of enzyme utilization. They enable enzymes to be reused after reactions, and in some rare cases increase enzymes activity (Bastida et al., 1998). Among the immobilization methods, for instance adsorption, entrapment (encapsulation), cross-linking and covalent bonds, adsorption has been the most widely used technique. Besides being practical, it causes less deterioration to enzyme activity. In some cases, adsorption can also combined with other methods (Aliyah et al., 2016). Similar to adsorption, entrapment in a resin matrix is a technique with a minimum deterioration effect. Although immobilization increases the reusability of enzymes, it frequently decreases their activity. Considering these possibilities, it is important to evaluate the application of both immobilized and free enzymes.
Frangipani (Plumeria rubra) latex is a source of lipase which displays remarkable activity (Cambon et al., 2006). Our previous work shows that the lipolytic activity of frangipani latex is found in the particulate fraction. However, its solid particulates are easily dissolved in an oil-water mixture and cannot be retained after a lipolysis reaction. Immobilization, despite involving more process steps, might reduce operational costs by the recycling of frangipani latex lipase. On the other hand, non-immobilized lipase is easier to prepare, although it is only available for single use. To develop a lipolysis technology utilizing frangipani latex lipase, it is important to evaluate its performance, both in immobilized and free (non-immobilized) form.
The aim of this study is to
compare the performance of immobilized lipase and free lipase from frangipani (Plumeria rubra) latex particulates. The
feasibility of such immobilization is evaluated in the study, and the effect of
denaturation is expected to be minimal. Immobilization was conducted with
methods that are less susceptible to enzyme denaturation, namely adsorption and
encapsulation (or entrapment). Frangipani latex lipase, in free or immobilized
form, could be a potential biocatalyst for fatty acid production with low
capital and operational costs, easy handling, and applicability in rural areas.
An effective method for immobilizing latex lipase will also be applicable for
lipase in solid form, such as dry extract lipase from microorganisms
(Hermansyah et al., 2018).
Immobilization methods for frangipani crude latex have been compared. Adsorption of particulate latex lipase was ineffective, although it works on liquid microbial lipase. Immobilization was successfully achieved with encapsulation in a calcium alginate matrix, producing a lower, yet homogenous, degree of lipolysis. This enables recycling of lipase, although with limited frequency, and it is also susceptible to contamination from broken matrix. The performance of free and immobilized lipases has also been evaluated. Free lipases produce a higher, yet heterogenous, degree of lipolysis than immobilized lipases. They are more suitable for small-scale lipolysis with a stirred tank to produce technical grade fatty acid. Meanwhile, immobilization of latex lipase requires further modification, such as use of a packed bed reactor, circulated flow, or matrix modification.
The authors are grateful to the Faculty of Industrial Technology, Institut Teknologi Bandung, for publication funding via a research grant awarded through the Research, Community Service and Innovation Program 2018 scheme, with Contract No. 0851b/I1.C06.2/PL/2018.
Abdelkafi S., Barouh N., Fouquet B., Fendri I., Pina M., Scheirlinckx F., Villeneuve P., Carrière F., 2011. Carica Papaya Lipase: A Naturally Immobilized Enzyme with Interesting Biochemical Properties. Plant Foods for Human Nutrition, Volume 66(1), pp. 34–40
Aliyah, A.N., Edelweiss, E.D., Sahlan, M., Wijanarko, A., Hermansyah, H., 2016. Solid State Fermentation using Agroindustrial Wastes to Produce Aspergillus Niger Lipase as a Biocatalyst Immobilized by an Adsorption-crosslinking Method for Biodiesel Synthesis. International Journal of Technology, Volume 7(8), pp. 1393–1404
Barnebey, H.L., Brown, A.C., 1948. Continuous Fat Splitting Plants using the Colgate-Emery Process. Journal of the American Oil Chemists’ Society, Volume 25(3), pp. 95–99
Barros, M., Fleuri, L.F., Macedo, G.A., 2010. Seed Lipases: Sources, Applications and Properties - A Review. Brazilian Journal of Chemical Engineering, Volume 27(1), pp. 15–29
Bastida, A., Sabuquillo, P., Armisen, P., Fernández-Lafuente, R., Huguet, J., Guisán, J.M., 1998. A Single Step Purification, Immobilization, and Hyperactivation of Lipases via Interfacial Adsorption on Strongly Hydrophobic Supports. Biotechnology and Bioengineering, Volume 58(5), pp. 486–493
Cambon, E., Gouzou, F., Pina, M., Barea, B., Barouh, N., Lago, R., Ruales, J., Tsai, S.W., Villeneuve, P., 2006. Comparison of the Lipase Activity in Hydrolysis and Acyl Transfer Reactions of Two Latex Plant Extracts from Babaco (Vasconcellea × Heilbornii Cv.) and Plumeria Rubra: Effect of the Aqueous Microenvironment. Journal of Agricultural and Food Chemistry, Volume 54(7), pp. 2726–2731
Hermansyah, H., Maresya, A., Putri, D.N., Sahlan, M., Meyer, M., 2018. Production of Dry Extract Lipase from Pseudomonas Aeruginosa by the Submerged Fermentation Method in Palm Oil Mill Effluent. International Journal of Technology, Volume 9(2), pp. 325–334
Istyami, A.N., Kresnowati, M.T.A.P., Prakoso, T., Soerawidjaja, T.H., 2018. The Use of Hydrophobic Beads in Triglyceride Hydrolysis. Journal of Advances in Technology and Engineering Research, Volume 4(1), pp. 9–16
Jin, Q., Jia, G., Zhang, Y., Yang, Q., Li, C., 2011. Hydrophobic Surface Induced Activation of Pseudomonas cepacia Lipase Immobilized into Mesoporous Silica. Langmuir, Volume 27(19), pp. 12016–12024
Manoel, E.A., Dos Santos, J.C.S., Freire, D.M.G., Rueda, N., Fernandez-Lafuente, R., 2015. Immobilization of Lipases on Hydrophobic Supports Involves the Open Form of the Enzyme. Enzyme and Microbial Technology, Volume 71, pp. 53–57
Mazou, M., Djossou, A.J., Tchobo, F.P., Villeneuve, P., Soumanou, M.M., 2016. Plant Latex Lipase as Biocatalysts for Biodiesel Production, African. Journal of Biotechnology, Volume 15(28), pp. 1487–1502
Mounguengui, R.W.M., Brunschwig, C., Barea, B., Villeneuve, P., Blin, J., 2013. Are Plant Lipases a Promising Alternative to Catalyze Transesterification for Biodiesel Production? Progress in Energy and Combustion Science, Volume 39(5), pp. 441–456
Rooney, D., Weatherley, L.R., 2001. The Effect of Reaction Conditions upon Lipase Catalysed Hydrolysis of High Oleate Sunflower Oil in a Stirred Liquid–liquid Reactor. Process Biochemistry, Volume 36(10), pp. 947–953
Seth, S., Chakravorty, D., Patra, S., 2014. An Insight into Plant Lipase Research - Challenges Encountered. Protein Expression and Purification, Volume 95, pp. 13–21