Displaying publications 1 - 20 of 84 in total

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  1. Leong HY, Chang CK, Khoo KS, Chew KW, Chia SR, Lim JW, et al.
    Biotechnol Biofuels, 2021 Apr 07;14(1):87.
    PMID: 33827663 DOI: 10.1186/s13068-021-01939-5
    Global issues such as environmental problems and food security are currently of concern to all of us. Circular bioeconomy is a promising approach towards resolving these global issues. The production of bioenergy and biomaterials can sustain the energy-environment nexus as well as substitute the devoid of petroleum as the production feedstock, thereby contributing to a cleaner and low carbon environment. In addition, assimilation of waste into bioprocesses for the production of useful products and metabolites lead towards a sustainable circular bioeconomy. This review aims to highlight the waste biorefinery as a sustainable bio-based circular economy, and, therefore, promoting a greener environment. Several case studies on the bioprocesses utilising waste for biopolymers and bio-lipids production as well as bioprocesses incorporated with wastewater treatment are well discussed. The strategy of waste biorefinery integrated with circular bioeconomy in the perspectives of unravelling the global issues can help to tackle carbon management and greenhouse gas emissions. A waste biorefinery-circular bioeconomy strategy represents a low carbon economy by reducing greenhouse gases footprint, and holds great prospects for a sustainable and greener world.
  2. Soh EYS, Lim SS, Chew KW, Phuang XW, Ho VMV, Chu KYH, et al.
    Environ Res, 2021 Nov 13.
    PMID: 34780790 DOI: 10.1016/j.envres.2021.112385
    The effluent of textile industries containing synthetic dyes contributed to substantial pollution to water bodies. The biosorption process of Congo Red dye was successfully performed by integrating ultrasonication in the adsorption step with spent brewery yeast as a novel and renewable biosorbent. The adsorption process was hindered when ultrasonication was employed together with the biosorbent, indicating that desorption process had occurred. The adsorption process showed that 4 g/L of biosorbent was the optimum dosage for adsorption of 50 mg/L of Congo Red dye, and that the adsorption equilibrium fitted to the Langmuir model, with kinetics best fitted with pseudo-second order model. The maximum capacity of the adsorption was 52.6 mg/g, showing the potential of spent brewery yeast to aid in removing wastewater pollutants. Maximal Congo Red dye recovery (100%) was achieved in the sonication-assisted desorption studies using 0.01M NaOH as the eluting agent. The ultrasonication effects contributed to the efficient recovery of dye and good conversion of spent brewery yeast to biosorbent can be beneficial for treating pollution from textile wastewater.
  3. Munawaroh HSH, Gumilar GG, Khoiriah SF, Nindya FS, Berliana N, Aisyah S, et al.
    PMID: 37363337 DOI: 10.1007/s12649-023-02141-4
    One of potential inhibitors which is widely used for the clinical treatment of COVID-19 in comorbid patients is Angiostensin Converting Enzyme-1 (ACE1) inhibitor. A safer peptide-based ACE1 inhibitor derived from salmon skin collagen, that is considered as the by-product of the fish processing industry have been investigated in this study. The inhibitory activity against ACE1 was examined using in vitro and in silico methods. In vitro analysis includes the extraction of acid-soluble collagen, characterization using FTIR, Raman, UV-Vis, XRD, cytotoxicity assay, and determination of inhibition against ACE1. In silico method visualizes binding affinity, molecular interaction, and inhibition type of intact collagen and active peptides derived from collagen against ACE1 using molecular docking. The results of FTIR spectra detected amide functional groups (A, B, I, II, III) and imine proline/hydroxyproline, while the results of Raman displayed peak absorption of amide I, amide III, proline/hydroxyproline ring, phenylalanine, and protein backbone. Furthermore, UV-Vis spectra showed typical collagen absorption at 230 nm and based on XRD data, the chain types in the samples were α-helix. ACE1 inhibition activity was obtained in a concentration-dependent manner where the highest was 82.83% and 85.84% at concentrations of 1000, and 2000 µg/mL, respectively, and showed very low cytotoxicity at the concentration less than 1000 µg/mL. In silico study showed an interaction between ACE1 and collagen outside the active site with the affinity of - 213.89 kcal/mol. Furthermore, the active peptides of collagen displayed greater affinity compared to lisinopril, namely HF (His-Phe), WYT (Trp-Tyr-Thr), and WF (Trp-Phe) of - 11.52; - 10.22; - 9.58 kcal/mol, respectively. The salmon skin-derived collagen demonstrated ACE1 inhibition activity with a non-competitive inhibition mechanism. In contrast, the active peptides were predicted as potent competitive inhibitors against ACE1. This study indicated that valorization of fish by-product can lead to the production of a promising bioactive compound to treat COVID-19 patient with diabetic comorbid.
  4. Pocha CKR, Chia SR, Chia WY, Koyande AK, Nomanbhay S, Chew KW
    Chemosphere, 2022 Mar;290:133246.
    PMID: 34906526 DOI: 10.1016/j.chemosphere.2021.133246
    The ever-growing human population has resulted in the expansion of agricultural activity; evident by the deforestation of rainfoamrests as a means of acquiring fertile land for crops. The crops and fruits produced by such means should be utilized completely; however, there are still losses and under-exploitation of these produces which has resulted in wastes being mounted in landfills. These underutilized agricultural wastes including vegetables and fruits can serve as a potential source for biofuels and green diesel. This paper discusses the main routes (e.g., biological and thermochemical) for producing biofuels such as bioethanol, biodiesel, biogas, bio-oil and green diesel from underutilized crops by emphasizing recent technological innovations for improving biofuels and green diesel yields. The future prospects of a successful production of biofuels and green diesel by this source are also explained. Underutilized lignocelluloses including fruits and vegetables serve as a prospective biofuel and green diesel generation source for the future prosperity of the biofuel industry.
  5. Tang DYY, Chew KW, Chia SR, Ting HY, Sia YH, Gentili FG, et al.
    Environ Technol, 2022 Dec 19.
    PMID: 36536589 DOI: 10.1080/09593330.2022.2150094
    Overgrowth of microalgae will result in harmful algae blooms that can affect the aquatic ecosystem and human health. Therefore, the quantitation of chlorophyll pigments can be used as an indicator of algae bloom. However, it is difficult to monitor the geographical and temporal distribution of chlorophyll in the aquatic environment. Accordingly, an innovative and inexpensive method based on the red-green-blue (RGB) image analysis was utilized in this study to estimate the microalgae chlorophyll content. The digital images were acquired using a smartphone camera. The colour index was then evaluated using software and associated with chlorophyll concentration significantly. A regression model, using RGB colour components as independent variables to estimate chlorophyll concentration, was developed and validated. The Green in the RGB index was the most promising way to estimate chlorophyll concentration in microalgae. The result showed that acetone was the best extractant solvent with a high R-squared value among the four extractant solvents. Next, the isolation of useful biomolecules, such as proteins, fatty acids, polysaccharides and antioxidants from the microalgae, has been recognized as an alternative to regulating algae bloom. Microalgae are shown to produce bioactive compounds with a variety of biological activities that can be applied in various industries. This study evaluates the biochemical composition of mixed microalgae species, Desmodesmus sp. and Scenedesmus sp. using the liquid triphasic partitioning (TPP) system. The findings from analytical assays revealed that the biomass consisted of varied concentrations of carbohydrates, protein, and lipids. Phenolic compounds and antioxidant activity were at 60.22 mg/L and 90.69%, respectively.
  6. Chong JWR, Khoo KS, Chew KW, Ting HY, Show PL
    Biotechnol Adv, 2023;63:108095.
    PMID: 36608745 DOI: 10.1016/j.biotechadv.2023.108095
    Identification of microalgae species is of importance due to the uprising of harmful algae blooms affecting both the aquatic habitat and human health. Despite this occurence, microalgae have been identified as a green biomass and alternative source due to its promising bioactive compounds accumulation that play a significant role in many industrial applications. Recently, microalgae species identification has been conducted through DNA analysis and various microscopy techniques such as light, scanning electron, transmission electron, and atomic force -microscopy. The aforementioned procedures have encouraged researchers to consider alternate ways due to limitations such as costly validation, requiring skilled taxonomists, prolonged analysis, and low accuracy. This review highlights the potential innovations in digital microscopy with the incorporation of both hardware and software that can produce a reliable recognition, detection, enumeration, and real-time acquisition of microalgae species. Several steps such as image acquisition, processing, feature extraction, and selection are discussed, for the purpose of generating high image quality by removing unwanted artifacts and noise from the background. These steps of identification of microalgae species is performed by reliable image classification through machine learning as well as deep learning algorithms such as artificial neural networks, support vector machines, and convolutional neural networks. Overall, this review provides comprehensive insights into numerous possibilities of microalgae image identification, image pre-processing, and machine learning techniques to address the challenges in developing a robust digital classification tool for the future.
  7. Foo WH, Chia WY, Tang DYY, Koay SSN, Lim SS, Chew KW
    J Hazard Mater, 2021 Sep 05;417:126129.
    PMID: 34229396 DOI: 10.1016/j.jhazmat.2021.126129
    Waste cooking oil (WCO) is considered as one of the hazardous wastes because improper disposal of WCO can cause significant environmental problems such as blockages of drains and sewers as well as water or soil pollution. In this review, the physical and chemical properties of WCO are evaluated along with its regulations and policies in different countries to promote WCO refined biofuels. Blended WCO can be an auxiliary fuel for municipal solid waste incinerators while the heat produced is able to form superheated steam and subsequently generate electricity via combined heat and power system. Also, WCO contains high ratio of hydrogen atoms compared to carbon and oxygen atoms, making it able to be catalytically cracked, synthesizing hydrogen gas. WCO-based biodiesel has been traditionally produced by transesterification in order to substitute petroleum-based diesel which has non-degradability as well as non-renewable features. Hence, the potentials of hazardous WCO as a green alternative energy source for electricity generation, hydrogen gas as well as biofuels production (e.g. biodiesel, biogas, biojet fuel) are critically discussed due to its attractive psychochemical properties as well as its economic feasibility. Challenges of the WCO utilization as a source of energy are also reported while highlighting its future prospects.
  8. Munawaroh HSH, Pratiwi RN, Gumilar GG, Aisyah S, Rohilah S, Nurjanah A, et al.
    Int J Biol Macromol, 2023 Mar 15;231:123248.
    PMID: 36642356 DOI: 10.1016/j.ijbiomac.2023.123248
    Gelatin hydrogel is widely employed in various fields, however, commercially available gelatin hydrogels are mostly derived from mammalian which has many disadvantages due to the supply and ethical issues. In this study, the properties of hydrogels from fish-derived collagen fabricated with varying Glutaraldehyde (GA) determined. The antidiabetic properties of salmon gelatin (SG) and tilapia gelatin (TG) was also evaluated against α-glucosidase. Glutaraldehyde-crosslinked salmon gelatin and tilapia gelatin were used, and compared with different concentrations of GA by 0.05 %, 0.1 %, and 0.15 %. Water absorbency, swelling, porosity, pore size and water retention of the hydrogels were dependent on the degree of crosslinking. The synthesis of hydrogels was confirmed by FTIR study. Scanning electron microscope (SEM) observation showed that all hydrogels have a porous structure with irregular shapes and heterogeneous morphology. Performance tests showed that gelatin-GA 0.05 % mixture had the best performance. Antidiabetic bioactivity in vitro and in silico tests showed that the active peptides of SG and TG showed a high binding affinity to α-glucosidase enzyme. In conclusion, SG and TG cross-linked GA 0.05 % have the potential as an antidiabetic agent and as a useful option over mammalian-derived gelatin.
  9. Chia WY, Chew KW, Le CF, Lam SS, Chee CSC, Ooi MSL, et al.
    Environ Pollut, 2020 Dec;267:115662.
    PMID: 33254731 DOI: 10.1016/j.envpol.2020.115662
    Acceleration of urbanization and industrialization has resulted in the drastic rise of waste generation with majority of them being biowaste. This constitutes a global challenge since conventional waste management methods (i.e., landfills) present environmental issues including greenhouse gases emissions, leachate formation and toxins release. A sustainable and effective approach to treat biowaste is through composting. Various aspects of composting such as compost quality, composting systems and compost pelletization are summarized in this paper. Common application of compost as fertilizer or soil amendment is presented with focus on the low adoption level of organic waste compost in reality. Rarely known, compost which is easily combustible can be utilized to generate electricity. With the analysis on critical approaches, this review aims to provide a comprehensive study on energy content of compost pellets, which has never been reviewed before. Environmental impacts and future prospects are also highlighted to provide further insights on application of this technology to close the loop of circular bioeconomy.
  10. Tham PE, Ng YJ, Vadivelu N, Lim HR, Khoo KS, Chew KW, et al.
    Bioresour Technol, 2022 Feb;346:126558.
    PMID: 34906702 DOI: 10.1016/j.biortech.2021.126558
    This research work aims to fabricate an optimized up-scaled photobioreactor and extraction tank which incorporates the Internet of Things (IoT) for remote monitoring of selected parameters without being present in the lab as the industry is gradually moving towards the direction of remote operation. Several design factors were considered where modelling using ANSYS was carried out before the finalised design is drawn using AutoCAD. To monitor critical parameters that include liquid level, temperature, and pH condition during the operation of the tanks, water-proof sensors are implemented with the aid of Arduino NodeMCU board and the sensors are linked with Blynk, a smartphone application that allows remote monitoring via Wi-Fi connection. The sensors' results obtained using the Blynk application show high accuracy as compared with manual data except for photobioreactor liquid level. This shows that IoT and remote monitoring can be integrated successfully.
  11. Chia SR, Chew KW, Show PL, Xia A, Ho SH, Lim JW
    Bioresour Technol, 2019 Oct;289:121727.
    PMID: 31279318 DOI: 10.1016/j.biortech.2019.121727
    In this present study, microalgal phycobiliproteins were isolated and purified via potential biphasic processing technique for pharmaceutical as well as food applications. The algal pre-treatment techniques were studied to enhance the yield of microalgal phycobiliproteins from the biomass. The proposed methods were optimised to obtain the best recovery yield of phycobiliproteins that can be isolated from the biomass. The phycobiliproteins were further purified using liquid biphasic system. The results showed that microalgal phycobiliproteins of high purity and yield was achieved using sonication treatment (20% power, 50% duty cycle and 7 min of irradiation time) with the biphasic system, where the purification fold of 6.17 and recovery yield of 94.89% was achieved. This work will provide insights towards the effective downstream processing of biomolecules from microalgae.
  12. Kurniawan TA, Haider A, Ahmad HM, Mohyuddin A, Umer Aslam HM, Nadeem S, et al.
    Chemosphere, 2023 Jun;325:138367.
    PMID: 36907482 DOI: 10.1016/j.chemosphere.2023.138367
    The generation of microplastics (MPs) has increased recently and become an emerging issue globally. Due to their long-term durability and capability of traveling between different habitats in air, water, and soil, MPs presence in freshwater ecosystem threatens the environment with respect to its quality, biotic life, and sustainability. Although many previous works have been undertaken on the MPs pollution in the marine system recently, none of the study has covered the scope of MPs pollution in the freshwater. To consolidate scattered knowledge in the literature body into one place, this work identifies the sources, fate, occurrence, transport pathways, and distribution of MPs pollution in the aquatic system with respect to their impacts on biotic life, degradation, and detection techniques. This article also discusses the environmental implications of MPs pollution in the freshwater ecosystems. Certain techniques for identifying MPs and their limitations in applications are presented. Through a literature survey of over 276 published articles (2000-2023), this study presents an overview of solutions to the MP pollution, while identifying research gaps in the body of knowledge for further work. It is conclusive from this review that the MPs exist in the freshwater due to an improper littering of plastic waste and its degradation into smaller particles. Approximately 15-51 trillion MP particles have accumulated in the oceans with their weight ranging between 93,000 and 236,000 metric ton (Mt), while about 19-23 Mt of plastic waste was released into rivers in 2016, which was projected to increase up to 53 Mt by 2030. A subsequent degradation of MPs in the aquatic environment results in the generation of NPs with size ranging from 1 to 1000 nm. It is expected that this work facilitates stakeholders to understand the multi-aspects of MPs pollution in the freshwater and recommends policy actions to implement sustainable solutions to this environmental problem.
  13. Neo YT, Chia WY, Lim SS, Ngan CL, Kurniawan TA, Chew KW
    Food Res Int, 2023 Mar;165:112480.
    PMID: 36869493 DOI: 10.1016/j.foodres.2023.112480
    Production and extraction systems of algal protein and handling process of functional food ingredients need to control several parameters such as temperature, pH, intensity, and turbidity. Many researchers have investigated the Internet of Things (IoT) approach for enhancing the yield of microalgae biomass and machine learning for identifying and classifying microalgae. However, there have been few specific studies on using IoT and artificial intelligence (AI) for production and extraction of algal protein as well as functional food ingredients processing. In order to improve the production of algal protein and functional food ingredients, the implementation of smart system is a must to have real-time monitoring, remote control system, quick response to sudden events, prediction and characterisation. Techniques of IoT and AI are expected to help functional food industries to have a big breakthrough in the future. Manufacturing and implementation of beneficial smart systems are important to provide convenience and to increase the efficiency of work by using the interconnectivity of IoT devices to have good capturing, processing, archiving, analyzing, and automation. This review investigates the possibilities of implementation of IoT and AI in production and extraction of algal protein and processing of functional food ingredients.
  14. Ullah H, Qureshi KS, Khan U, Zaffar M, Yang YJ, Rabat NE, et al.
    Chemosphere, 2021 Dec;285:131492.
    PMID: 34273691 DOI: 10.1016/j.chemosphere.2021.131492
    The restoration of mechanical properties is desired for creating the self-healing coatings with no corrosion capabilities. The encapsulation of epoxy resins is limited by various factors in urea and melamine formaldehyde microcapsules. An improved method was developed, where epoxy resin was encapsulated by individual wrapping of poly(melamine-formaldehyde) and poly(urea-formaldehyde) shell around emulsified epoxy droplets via oil-in-water emulsion polymerization method. The synthesized materials were characterized analytically. The curing of the epoxy was achieved by adding the [Ni/Co(2-MI)6].2NO3 as a latent hardener and iron acetylacetonate [Fe(acac)3] as a latent accelerator. Isothermal and non-isothermal differential scanning calorimetric analysis revealed lower curing temperature (Tonset = 116 °C) and lower activation energies (Ea ≈ 69-75 kJ/mol). The addition of microcapsules and complexes did not adversely alter the flexural strength and flexural modulus of the epoxy coatings. The adhesion strength of neat coating decreased from 6310.8 ± 31 to 4720.9 ± 60 kPa and percent healing increased from 50.83 to 67.45% in the presence of acetylacetonate complex at 10 wt% of microcapsules.
  15. Leong YK, Chew KW, Chen WH, Chang JS, Show PL
    Trends Plant Sci, 2021 07;26(7):729-740.
    PMID: 33461869 DOI: 10.1016/j.tplants.2020.12.010
    Given their advantages of high photosynthetic efficiency and non-competition with land-based crops, algae, that are carbon-hungry and sunlight-driven microbial factories, are a promising solution to resolve energy crisis, food security, and pollution problems. The ability to recycle nutrient and CO2 fixation from waste sources makes algae a valuable feedstock for biofuels, food and feeds, biochemicals, and biomaterials. Innovative technologies such as the bicarbonate-based integrated carbon capture and algae production system (BICCAPS), integrated algal bioenergy carbon capture and storage (BECCS), as well as ocean macroalgal afforestation (OMA), can be used to realize a low-carbon algal bioeconomy. We review how algae can be applied in the framework of integrated low-carbon circular bioeconomy models, focusing on sustainable biofuels, low-carbon feedstocks, carbon capture, and advances in algal biotechnology.
  16. Ali S, Paul Peter A, Chew KW, Munawaroh HSH, Show PL
    Bioresour Technol, 2021 Jun 26;337:125461.
    PMID: 34198241 DOI: 10.1016/j.biortech.2021.125461
    Industrial effluents such as pharmaceutical residues, pesticides, dyes, and metal processes holds abundant value-added products (VAPs), where its recovery has become essential. The purpose of such recovery is for sustainable treatment, which is an approach that considers the economic, social, and environmental aspects. Microalgae with its potential in the recovery process from effluents, can reduce energy usage of waste management strategies and regenerate nutrients such as carbon, phosphorus, and nitrogen. Microalgae cultures offer the use of inorganic materials by microalgae for their growth and the help of bacteria to produce biomass, thus, resulting in the absence of secondary emissions due to its ability to eliminate volatile organic compounds. Moreover, recovered bioactive compounds are transformed into bioethanol, bio-fertilizers, biopolymer, health supplements and animal feed. Therefore, it is significant to focus on an economical and efficient utilization of microalgae in recovering nutrients that can be further used in various commercial applications.
  17. Khoo KS, Chew KW, Yew GY, Leong WH, Chai YH, Show PL, et al.
    Bioresour Technol, 2020 May;304:122996.
    PMID: 32115347 DOI: 10.1016/j.biortech.2020.122996
    The world energy system faces two major challenges: the requirement for more energy and less carbon. It is important to address biofuels production as an alternative to the usage of fossil fuel by utilizing microalgae as the potential feedstock. Yet, the commercialization of microalgae remains contentious caused by factors relating to the life cycle assessment and feasibility of microalgae-based biofuels. This present review starts with an introduction to the benefits of microalgae, followed by intensive elaboration on microalgae cultivation parameters. Subsequently, the fundamental principle along with the advantages and disadvantages of various pretreatment techniques of microalgae were reviewed. In addition, the conventional and recent advances in lipid extraction techniques from microalgae were comprehensively evaluated. Comparative analysis regard to the gaps from previous studies was discussed point-by-point in each section. The effort presented in this review will provide an insight for future researches dealing with microalgae-biofuel production on downstream processing.
  18. Chan SS, Khoo KS, Chew KW, Ling TC, Show PL
    Bioresour Technol, 2022 Jan;344(Pt A):126159.
    PMID: 34673198 DOI: 10.1016/j.biortech.2021.126159
    The litter of persistent organic pollutants (POPs) into the water streams and soil bodies via industrial effluents led to several adverse effects on the environment, health, and ecosystem. For the past decades, scientists have been paying efforts in the innovation and development of POPs removal from wastewater treatment. However, the conventional methods used for the removal of POPs from wastewater are costly and could lead to secondary pollution including soil and water bodies pollution. In recent, the utilization of green mechanisms such as biosorption, bioaccumulation and biodegradation has drawn attention and prelude the potential of green technology globally. Microalgae-bacteria consortia have emerged to be one of the latent wastewater treatment systems. The synergistic interactions between microalgae and bacteria could proficiently enhance the existing biological wastewater treatment system. This paper will critically review the comparison of conventional and recent advanced wastewater treatment systems and the mechanisms of the microalgae-bacteria symbiosis system.
  19. Ng YJ, Chan SS, Khoo KS, Munawaroh HSH, Lim HR, Chew KW, et al.
    Biotechnol Adv, 2023 Nov;68:108198.
    PMID: 37330152 DOI: 10.1016/j.biotechadv.2023.108198
    Surfactants have always been a prominent chemical that is useful in various sectors (e.g., cleaning agent production industry, textile industry and painting industry). This is due to the special ability of surfactants to reduce surface tension between two fluid surfaces (e.g., water and oil). However, the current society has long omitted the harmful effects of petroleum-based surfactants (e.g., health issues towards humans and reducing cleaning ability of water bodies) due to their usefulness in reducing surface tension. These harmful effects will significantly damage the environment and negatively affect human health. As such, there is an urgency to secure environmentally friendly alternatives such as glycolipids to reduce the effects of these synthetic surfactants. Glycolipids is a biomolecule that shares similar properties with surfactants that are naturally synthesized in the cell of living organisms, glycolipids are amphiphilic in nature and can form micelles when glycolipid molecules clump together, reducing surface tension between two surfaces as how a surfactant molecule is able to achieve. This review paper aims to provide a comprehensive study on the recent advances in bacteria cultivation for glycolipids production and current lab scale applications of glycolipids (e.g., medical and waste bioremediation). Studies have proven that glycolipids are effective anti-microbial agents, subsequently leading to an excellent anti-biofilm forming agent. Heavy metal and hydrocarbon contaminated soil can also be bioremediated via the use of glycolipids. The major hurdle in the commercialization of glycolipid production is that the cultivation stage and downstream extraction stage of the glycolipid production process induces a very high operating cost. This review provides several solutions to overcome this issue for glycolipid production for the commercialization of glycolipids (e.g., developing new cultivating and extraction techniques, using waste as cultivation medium for microbes and identifying new strains for glycolipid production). The contribution of this review aims to serve as a future guideline for researchers that are dealing with glycolipid biosurfactants by providing an in-depth review on the recent advances of glycolipid biosurfactants. By summarizing the points discussed as above, it is recommended that glycolipids can substitute synthetic surfactants as an environmentally friendly alternative.
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