Utilizing rice straw and sugarcane bagasse as low-cost feedstocks towards sustainable production of succinic acid.

Succinic acid (SA) has been produced from rice straw (RS) and sugarcane bagasse (SB) as low-cost feedstocks in this study through sequential peracetic acid (PA) and alkaline peroxide (AP) pretreatment assisted by ultrasound and pre-hydrolysis followed by simultaneous saccharification and fermentation (PSSF). The effect of yeast extract (YE) concentration, inoculum concentration, and biomass type on SA production was investigated. The results showed that SA production from RS and SB was significantly affected by the YE concentration. Final concentration and yield of SA produced were significantly increased along with the increasing of YE concentration. Moreover, inoculum concentration significantly affected the SA production from SB. Higher inoculum concentration led to higher SA production. On the other hand, SA production from RS was not significantly affected by the inoculum concentration. Using RS as the feedstock, the highest SA production was achieved on the medium containing 15 g/L YE with 5 % v/v inoculum, obtaining SA concentration and yield of 3.64 ± 0.1 g/L and 0.18 ± 0.05 g/g biomass, respectively. Meanwhile, the highest SA production from SB was acquired on the medium containing 10 g/L YE with 7.5 % v/v inoculum, resulting SA concentration and yield of 5.1 ± 0.1 g/L and 0.25 ± 0.05 g/g biomass, respectively. This study suggested that RS and SB are potential to be used as low-cost feedstocks for sustainable and environmentally friendly SA production through ultrasonic-assisted PA and AP pretreatment and PSSF.

Optimization of Ultrasonic-Assisted Extraction (UAE) Method Using Natural Deep Eutectic Solvent (NADES) to Increase Curcuminoid Yield from Curcuma longa L., Curcuma xanthorrhiza, and Curcuma mangga Val.

This study aims to optimize ultrasonic-assisted natural deep eutectic solvents (NADES) based extraction from C. longa. Choline chloride-lactic acid (CCLA-H2O = 1:1, b/v) was used to investigate the impact of various process parameters such as solvent's water content, solid loading, temperature, and extraction time. The optimal yield of 79.635 mg/g of C. longa was achieved from extraction in 20% water content NADES with a 4% solid loading in 35 °C temperature for 1 h. Peleg's model was used to describe the kinetics of the optimized ultrasonic-assisted extraction (UAE) method, and the results were found to be compatible with experimental data. The optimum conditions obtained from C. longa extraction were then used for the extraction of C. xanthorriza and C. mangga, which give yields of 2.056 and 31.322 mg/g, respectively. Furthermore, n-hexane was utilized as an anti-solvent in the separation process of curcuminoids extract from C. longa, C. xanthorriza, and C. mangga, which gave curcuminoid recovery of 39%, 0.74%, and 27%, respectively. Solidification of curcuminoids was also carried out using the crystallization method with n-hexane and isopropanol. However, the solution of CCLA and curcuminoids formed a homogeneous mixture with isopropanol. Hence, the curcuminoids could not be solidified due to the presence of NADES in the extract solution.

Towards efficient and greener processes for furfural production from biomass: A review of the recent trends.

As mentioned in several recent reviews, biomass-based furfural is attracting increasing interest as a feasible alternative for the synthesis of a wide range of non-petroleum-derived compounds. However, the lack of environmentally friendly, cost-effective, and sustainable industrial procedures is still evident. This review describes the chemical and biological routes for furfural production. The mechanisms proposed for the chemical transformation of xylose to furfural are detailed, as are the current advances in the manufacture of furfural from biomass. The main goal is to overview the different ways of improving the furfural synthesis process. A pretreatment process, particularly chemical and physico-chemical, enhances the digestibility of biomass, leading to the production of >70 % of available sugars for the production of valuable products. The combination of heterogeneous (zeolite and polymeric solid) catalyst and biphasic solvent system (water/GVL and water/CPME) is regarded as an attractive approach, affording >75 % furfural yield for over 80 % of selectivity with the possibility of catalyst reuse. Microwave heating as an activation technique reduces reaction time at least tenfold, making the process more sustainable. The state of the art in industrial processes is also discussed. It shows that, when sulfuric acid is used, the furfural yields do not exceed 55 % for temperatures close to 180 °C. However, the MTC process recently achieved an 83 % yield by continuously removing furfural from the liquid phase. Finally, the CIMV process, using a formic acid/acetic acid mixture, has been developed. The economic aspects of furfural production are then addressed. Future research will be needed to investigate scaling-up and biological techniques that produce acceptable yields and productivities to become commercially viable and competitive in furfural production from biomass.

Fluoride and calcium release from peppermint-flavored fluoride varnish containing dicalcium-phosphate-dihydrate coated with xylitol.

Bakground: Fluoride varnish with high initial fluoride and calcium release can help patients with high-risk caries. Ample quantities of free fluoride and calcium ions in the oral cavity can enhance enamel remineralization. This study aimed to investigate the effect of dicalcium phosphate dihydrate coated with xylitol (DCPD-xylitol), in fluoride varnish, on the release of fluoride and calcium ions in the oral cavity. MATERIALS AND METHODS: DCPD powder with xylitol was synthesized by preparing a 60% xylitol solution and mixed it with DCPD solution. The mixture was stirred for 1 h at room temperature and dried at 80 °C for 18 h to reduce the water content. Then, the powder was used in the formulation of peppermint-flavored fluoride varnish as an active agent. The amounts of fluoride and calcium ion released in deionized water at 37 °C for 6 h were assessed with an ion-selective electrode. The cumulative fluoride and calcium ion release data were analyzed using one-way analysis of variance (ANOVA) and the post hoc Tukey test with α = 0.05. RESULTS: The results showed that the addition of DCPD coated with xylitol provided better bioavailability of the ions than DCPD without coating. Peppermint-flavored fluoride varnish (PFFV) with DCPD-xylitol 1% gave the highest fluoride ion release (296.90 mg/L) compared to the varnishes with other xylitol concentrations and the positive control. In contrast, PFFV DCPD-xylitol 5% afforded the highest calcium ion release at 111.20 mg/L. CONCLUSIONS: This study concluded that xylitol affects the bioavailability of free fluoride and calcium ions in varnishes. However, the efficacy of fluoride and calcium uptake in enamel and under different in vitro media conditions requires further investigation.

Immobilization of Cholesterol Oxidase from Streptomyces Sp. on Magnetite Silicon Dioxide by Crosslinking Method for Cholesterol Oxidation.

Enzymatic biosensor has been paid much attention to the research fields due to its advantage in medical application. As one of the application, we determined the optimum value of cholesterol oxidase against cholesterol. In this work, we studied the behavior of cholesterol oxidation by enzymatic reaction to get the optimum condition for cholesterol oxidation. The enzyme that used were in two form, free cholesterol oxidase, and immobilized cholesterol oxidase. Cholesterol oxidase was produced from Streptomyces sp. by using solid state fermentation method and identified had high enzyme activity to be 5.12 U/mL. Cholesterol oxidase was simultaneously crosslinked immobilized onto magnetite coated by silicon dioxide (M-SiO2). The support was characterized by Fourier transform infrared (FTIR) to determine the functional group of modified particle and scanning electron microscope (SEM) to observe the morphological or our prepared particle. Cholesterol oxidase sensitivity to substrate was analyzed by using HPLC with different interval time measurements. The oxidation of cholesterol by free enzyme and immobilized enzyme was also investigated. The best sensitivity of cholesterol oxidase was estimated to oxidize Cso (concentration of substrate) 1.46 mM of substrate with Ce (concentration of enzyme) 20 mg/mL for 180 min. Final oxidation value of cholesterol by immobilized enzyme was greater than 60%. The results of this study revealed that immobilized enzyme for cholesterol oxidation was stable, reproducible, and sensitive.

Purification and characterization of proteins in multifloral honey from kelulut bee (stingless bee).

The kelulut bee (Meliponini) is a subfamily of stingless bees that produce honey. A total of 89 species out of a total of 500 species of kelulut bees are known to originate from the Indo-Australian region. Kelulut bees do not have quality standards so they still refer to the Codex and EU Directive which basically only applied for Apis honey. The Codex and EU Directive are formed by several psychochemical parameters, one of it is diastase activity. Diastase activity in kelulut honey is known not to meet existing standards or even undetectable. Therefore, this study aimed to explore proteins inside kelulut honey and investigate the possibility of using a specific protein as a biomarker to differentiate honey produced by kelulut bee from other honey. This research can also be considered as an initial step to optimize the exploration of protein in kelulut honey. This research is divided into two sections which are the preliminary research and the research expansion. From preliminary section, glucose dehydrogenase enzyme (GDH) was found to be present inside Tetragonula spp honey. A further examination of GDH enzyme was made in four kelulut bee honeys namely Tetragonula leaviceps, T. biroi, Heterotrigona itama, and Geniotrigona thoracica. The preliminary research has five stages that are exactly the as expansion research section except it didn't include GDH activity measurement. The research includes seven main stages. First honeys were dialyzed to remove the sugar content followed by centrifugation. The samples were then purified using liquid chromatography with anion exchanger column. The molecular weight of proteins was analysed by SDS-PAGE method. The GDH activity was measured using spectrophotometer followed by qualitative analysis using LC-MS/MS. The peptide sequences resulted from LC-MS/MS were then matched with Uniprot to identify the unknow protein. The results showed that only T. biroi and T. laeviceps had GDH enzyme activity of 0,1891 U/mL and 0,1652-1,579 U/mL, respectively. Bands from both species were also qualitatively identified as GDH. With these results, it can be concluded that the GDH enzyme cannot be used as a biomarker to distinguish the kelulut honey.

Identification and classification of honey's authenticity by attenuated total reflectance Fourier-transform infrared spectroscopy and chemometric method.

BACKGROUND AND AIM: The authentication of honey is important to protect industry and consumers from such adulterated honey. However, until now, there has been no guarantee of honey's authenticity, especially in Indonesia. The classification of honey is based on the bee species (spp.) that produces it. The study used honey from sting bee Apis spp. and stingless bee Tetragonula spp. based on the fact that the content off honey produced between them has differences. Authenticating honey with currently available rapid detection methods, such as 13C nuclear magnetic resonance analysis, is costly. This study aimed to develop an inexpensive, fast, precise, and accurate classification method for authenticating honey. MATERIALS AND METHODS: In this study, we use attenuated total reflectance Fourier-transform infrared (ATR-FTIR) spectroscopy with wavelengths ranging between 550 and 4000 cm-1 as an alternative analysis method, which is relatively less expensive. The spectra of authentic and fake honey samples were obtained using ATR-FTIR and plotted using chemometric discriminant analysis. The authentic honey samples were acquired from a local Indonesian breeder of honey bees, while the fake honey samples were made from a mixture of water, sugar, sodium bicarbonate, and authentic honey. Data were collected using Thermo Scientific's OMNIC FTIR software and processed using Thermo Scientific's TQ Analyst software. RESULTS: Our method effectively classified the honey as authentic or fraudulent based on the FTIR spectra. To authenticate the honey, we formed two classes: Real honey and fake honey. The wavelengths that can best differentiate between these two classes correspond to four regions: 1600-1700 cm-1; 1175-1540 cm-1; 940-1175 cm-1; and 700-940 cm-1. Similarly, for classification purpose, we formed two classes: Apis spp. and Tetragonula spp. The wavelength region that can best classify the samples as belonging to the Apis spp. or Tetragonula spp. class is explicitly within the range of 1600-1700 cm-1. CONCLUSION: This study successfully demonstrated a method to rapidly and accurately classify and authenticate honey. ATR-FTIR is a useful tool to test the authenticity of honey.

Antiretroviral activity of Pterois volitans (red lionfish) venom in the early development of human immunodeficiency virus/acquired immunodeficiency syndrome antiretroviral alternative source.

AIM: This study aimed to investigate the antiviral activity of Pterois volitans phospholipase A2 (PV-PLA2) from Indonesia to human immunodeficiency virus (HIV). MATERIALS AND METHODS: Fresh venomous fin parts of wild PV specimens were collected from Java Sea waters. Then, it washed using phosphate buffer pH 7.0 and immersed in phosphate buffer pH 7.0 0.01 m containing CaCl2 0.001 m for 24 h. The immersed fin then allowed for extraction process by sonicating for 2×8 min with 80% pulse and 20 kHz output with temperature controlling to avoid denaturation. The crude venom (CV) extracted from the fin is allowed for purification by 80% ethanol (ET) precipitation and ammonium sulfate fractionation method. The purified PV-PLA2 then analyzed using Lowry's method, Marinette's method, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and 3-(4, 5-dimethyl thiazol-2yl)-2, 5-diphenyl tetrazolium bromide assay. After determining the purest and safest sample of six samples analyzed, the chosen sample then tested into simian retrovirus-2 (SRV2)-A549 culture (48×104 cells/mL at 1-4 ppm), and compared to the CV sample (1-4 ppm) and lamivudine (100 ppm). The culture then is analyzed using a quantitative real time-polymerase chain reaction to find out the copy number of SRV-2 virus in each culture. RESULTS: The protein's activity, concentration, and purity analysis revealed that the PV-PLA2 purified using ammonium sulfate fractionation has the highest activity (1.81 times higher than the CV at 80% fractionation) and has higher purity than the sample from ET fractionation. The testing of the sample purified using ammonium sulfate fractionation at 80% saturation level shown that it has a 97.78% inhibition level toward SRV2-A549 culture at 4 ppm. However, in comparison to lamivudine which has 99.55% inhibition level at 100 ppm, it needs much lower concentration to achieve the same result. CONCLUSION: The significant inhibition of SRV2-A549 culture shown that the PV-PLA2 extracted from PV venom has the potential to become anti-HIV substances. It would be worthwhile to further evaluate the antiretroviral activity of PV-PLA2 in the in vivo studies.

Antiviral activity of Acanthaster planci phospholipase A2 against human immunodeficiency virus.

AIM: Investigation of antiviral activity of Acanthaster planci phospholipase A2 (AP-PLA2) from moluccas to human immunodeficiency virus (HIV). MATERIALS AND METHODS: Crude venom (CV) and F20 (PLA2 with 20% fractioned by ammonium sulfate) as a sample of PLA 2 obtained from A. planci's extract were used. Enzymatic activity of PLA2 was determined using the degradation of phosphatidylcholine (PC). Activity test was performed using in vitro method using coculture of phytohemagglutinin-stimulated peripheral blood mononuclear cell (PBMC) from a blood donor and PBMC from HIV patient. Toxicity test of AP-PLA2 was done using lethal concentration required to kill 50% of the population (LC50). RESULTS: AP-PLA2 F20 had activity and purity by 15.66 times bigger than CV. The test showed that the LC50 of AP-PLA2 is 1.638 mg/ml. Antiviral analysis of AP-PLA2 in vitro showed the inhibition of HIV infection to PBMC. HIV culture with AP-PLA2 and without AP-PLA2 has shown the number of infected PBMC (0.299±0.212% and 9.718±0.802%). Subsequently, RNA amplification of HIV using reverse transcriptase-polymerase chain reaction resulted in the decrease of band intensity in gag gene of HIV. CONCLUSION: This research suggests that AP-PLA2 has the potential to develop as an antiviral agent because in vitro experiment showed its ability to decrease HIV infection in PBMC and the number of HIV ribonucleic acid in culture.

Rigorous kinetic model considering positional specificity of lipase for enzymatic stepwise hydrolysis of triolein in biphasic oil-water system.

A rigorous kinetic model describing the stepwise triglyceride hydrolysis at the oil-water interface, based on the Ping Pong Bi Bi mechanism using suspended lipase having positional specificity, was constructed. The preference of the enzyme to cleave to the ester bonds at the edge and the center of the glycerol backbone of the substrates (tri-, di- or monoglyceride) was incorporated in the model. This model was applied to the experimental results for triolein hydrolysis using suspended Porcine pancreatic lipase (an sn-1,3 specific lipase) and Candida rugosa lipase (a non-specific lipase) in a biphasic oil-water system under various operating conditions. In order to discuss the model's advantages, other models that do not consider the positional specificity of the lipase were also applied to our experimental results. The model considering the positional specificity of the lipase gave results which fit better with the experimental data and described the effect of the initial enzyme concentration, the interfacial area, and the initial concentrations of triolein on the entire process of the stepwise triolein hydrolysis. This model also gives a good representation of the rate for cleaving the respective ester bonds of each substrate by each type of lipase.