Purification Factor Research Articles

Enhanced laccase separation from fermentation medium using cryogel columns

The laccase enzyme family belongs to the oxidoreductase enzyme class and is one of the most commercially valuable enzymes that catalyzes the oxidation of one electron of a wide range of phenolic compounds. Separation and purification of laccases are crucial for industry since they play an important role in dye decolorization, biodegradation and food processing. Therefore, developing effective, high yielding and cost-effective methods for laccase production is vital. In this study, it was aimed to prepare cryogel columns for laccase purification following the bioproduction of laccase via Aspergillus niger. 2-hydroxyethyl methacrylate based cryogels were synthesized in the presence of 1-vinylimidazole as the affinity ligand and characterized by swelling tests, Brunauer–Emmett–Teller surface area measurement and scanning electron microscopy analysis. Surface area and water uptake ratio of cryogel columns were 35 m2/g and 93 %, respectively. The effect of pH, equilibrium laccase concentration, flow rate, interaction time and temperature on laccase adsorption were examined. The purification factor was calculated as 10.53 under optimum conditions and the enzyme recovery was found to be 86.7 % from fermentation medium. Current study revealed that laccase purification using cryogels following filtration of fermentation medium could be a promising candidate for industrial applications with eliminating the need for complex chromatographic steps.

Production, characterization and techno-economic evaluation of Aspergillus fusant l-asparaginase

Protoplast fusion is one of the most reliable methods of introducing desirable traits into industrially-promising fungal strains. It harnesses the entire genomic repertoire of fusing microorganisms by routing the natural barrier and genetic incompatibility between them. In the present study, the axenic culture of a thermo-halotolerant strain of Aspergillus candidus (Asp-C) produced an anti-leukemic l-asparaginase (l-ASNase) while a xylan-degrading strain of Aspergillus sydowii (Asp-S) produced the acrylamide-reduction type. Protoplast fusion of the wild strains generated Fusant-06 with improved anti-leukemic and acrylamide reduction potentials. Submerged fed-batch fermentation was preferred to batch and continuous modes on the basis of impressive techno-economics. Fusant-06 l-ASNase was purified by PEG/Na+ citrate aqueous two-phase system (ATPS) to 146.21-fold and global sensitivity analysis report revealed polymer molecular weight and citrate concentration as major determinants of yield and purification factor, respectively. The enzyme was characterized by molecular weight, amino acid profile, activity and stability to chemical agents. Michaelis–Menten kinetics, evaluated under optimum conditions gave Km, Vmax, Kcat, and Kcat/Km as 6.67 × 10–5 M, 1666.67 µmolmin−1 mg−1 protein, 3.88 × 104 min−1 and 5.81 × 108 M−1.min−1 respectively. In-vitro cytotoxicity of HL-60 cell lines by Fusant-06 l-ASNase improved significantly from their respective wild strains. Stability of Fusant-06 l-ASNase over a wide range of pH, temperature and NaCl concentration, coupled with its micromolar Km value, confers commercial and therapeutic value on the product. Free-radical scavenging and acrylamide reduction activities were intermediate and the conferred thermo-halo-stability could be exploited for sustainable clinical and food industry applications.

Purification of lipase from Burkholderia metallica fermentation broth in a column chromatography using polymer impregnated resins

In this work, porous glass beads grafted with polyethylene glycol (PEG) were used as an adsorbent to purify lipase from Burkholderia metallica in column chromatography. The purification parameters viz. salt stability, types and concentrations of PEG and salt, pH of the binding solution, and flow rate were studied to determine the performance of the purification system in an XK16/20 column. The crude lipase was mixed with different types and concentrations of salts 1–5% (w/w) (sodium citrate, potassium citrate, and sodium acetate) and subjected to the column containing the polymeric glass bead. One-variable-at-a-time experimentation revealed that 20% (w/w) PEG 6000 g/mol impregnated glass beads with a binding solution of 5% sodium citrate at pH 7.7, a flow rate of 1.0 mL/min and extraction time of 10 min resulted in the highest purification factor and recovery yield at 3.67 and 88%, respectively. The purified lipase has 55 ∼ 60 kDa molecular mass. The outcome of the study showed PEG could be applied to modify the inert glass beads into polymeric form, providing a biocompatible and mild separation condition for lipase. Thus, PEG could be successfully applied for the purification of lipase from B. metallica fermentation broth using column chromatography.

CAN ZAKAT AND PURIFICATION BE EMPLOYED IN PORTFOLIO MODELLING?

The Capital Asset Pricing Model (CAPM), which has interest rates in its specification, can be deemed non-Shariah compliant. Therefore, the sukuk rate has been proposed to replace these rates in CAPM. This study analyses portfolio modelling by involving two essential elements in Islamic principles, namely zakat and purification. The concept of purification has been applied in the Shariah stock selection process in Indonesia, while at the same time, zakat has been widely socialised in stock investment. The study highlights two models that consider the concept of zakat reduction and the purification factors for portfolios in the Indonesian stock market. According to the robustness tests conducted, the proposed Shariah-compliant asset pricing model developed in this study is valid. Zakat reduction and purification factor integration in mathematical models can be applied in portfolio modelling.

Investigation and Screening of Mixed Microalgae Species for Lipase Production and Recovery using Liquid Biphasic Flotation Approach

BackgroundLipase is mainly extracted from animals, plants, fungi, and bacteria. The difficulty in extracting lipase from these organisms has led to the discovery of sustainable sources producing lipase. This study is the first to document the existence of lipase in the microalgae species from Umeå. MethodsLipase in microalgae was extracted using liquid biphasic flotation as the protein homogenate from the extracts was found to contain lipase activity. The lipase activity was evaluated using colorimetric method with p-nitrophenyl palmitate as the substrate. The types, volumes, and concentrations of solvents as well as flotation kinetics, and the biomass weight were optimized. The experiment was conducted at optimal temperature and pH 7. Significant FindingsThe optimized conditions for maximum lipase production were 400 mg of biomass, 10 min flotation time with flotation rate of 100 cc/min as well as 99.8% ethanol and 300 g/L ammonium sulphate concentration with volume ratio of 1: 0.83. Lipase recovery yield of the optimal LBF system was 70.3% with the separation efficiency of 82.0% and purification factor of 7.45. The findings revealed that the microalgae can be an ideal candidate for producing lipase and LBF serves as the platform for the development of large-scale extraction system.

Synthesis and characterization of supermacroporous cryogel with immobilized p-aminobenzenesulfonamide as affinity ligand for the purification of lactoperoxidase from whey.

This study proposed the development of a monolithic supermacroporous affinity column for direct capture of lactoperoxidase, a glycoprotein present in milk, whey, and colostrum, with several applications due to its wide antimicrobial activity. A poly(acrylamide)-based cryogel was produced by radical co-polymerization of monomers in frozen aqueous solution and activated with p-aminobenzenesulfonamide as a ligand for specific interaction with the lactoperoxidase. The axial liquid dispersion coefficients at different liquid flow rates were determined by measuring residence time distributions using the tracer pulse-response method. The axial dispersion coefficient was low and the height equivalent to theoretical plate was not dependent on the flow velocity. The adsorptive capacity of affinity cryogel was studied as a function of flow velocity and the best condition was 0.9cm/min. The response surface methodology was applied to optimize the capture of the enzyme, as a function of pH and salt concentration. Higher purification factor value was found at a salt concentration of 80mmol/L and pH of 8.0 (p<0.05). There was no influence of the variables under study on the yield (p>0.05). The results indicated that affinity cryogel is a promising chromatography support for the use in high-throughput one-step purification of lactoperoxidase from whey.

S-scheme tubular g-C3N4/BiOI heterojunctions for boosting photodegradation of tetracycline and Cr(VI): Mechanism insight, degradation pathway and DFT calculation

A unique tubular g-C3N4 (TCN)/BiOI S-scheme heterojunction with BiOI microspheres grown on the surface of 1D TCN was successfully prepared by hydrothermal treatment and calcination. Herein, adjusting BiOI additive amount can effectively tune electronic band structures and photocatalytic properties of TCN/BiOI heterojunctions. Impressively, the TCN/BiOI composite (CB-2) with a TCN: Bi3+ molar ratio of 1:2 shows the best photocatalytic performance, where the photodegradation efficiencies for tetracycline (TC) and Cr(VI) ions can both achieve more than 77 % and 90 % within 10 and 30 min, respectively. Particularly, the photodegradation efficiencies for TC and Cr(VI) reach up to 90.1 % and 96.8 % at 30 and 40 min, respectively. Notably, the rate constants for photodegradation of TC and Cr(VI) reach 0.1687 and 0.0718 min−1, which are 27.2 and 8.5 times higher than those of bulk g-C3N4, respectively. The distinguished photocatalytic performance of CB-2 is attributed to the high-quality electron transport channels provided by its tubular structure and the effective separation of photogenerated carriers based on the S-scheme heterojunction. Further, O2−, 1O2 and h+ are manifested to be the main active species for the photodegradation of TC. DFT calculations and radical-related experiments fully confirm the formation of S-scheme heterojunctions. Several crucial factors (such as pH, inorganic salts) for wastewater purification are verified to be little effect on the photocatalytic performance. The degradation pathways of TC are proposed. This work establishes a new route to design and prepare high-efficiency S-scheme photocatalysts for the photodegradation of antibiotics and the photoreduction of heavy metal ions in wastewater.

Performance of pseudo‐specific cryogel in lysozyme purification from chicken egg white

AbstractThe application of cryogels for biomolecule purification has expanded due to their adsorption efficiency and operational advantages. In this study, polyacrylamide cryogels functionalized with l‐phenylalanine (cryogel‐Phe) via the glutaraldehyde method were designed for lysozyme adsorption. Cryogel functionalization was confirmed by Fourier‐transform infrared spectroscopy and Kjeldahl analysis, indicating the immobilization of 458.65 mgphenylalanine gcryogel−1. Cryogel‐Phe showed high porosity (0.95) and a Young's modulus of 526.71 kPa. Thermogravimetric analysis indicated that thermal degradation occurred above 200°C. Differential scanning calorimetry and X‐ray diffraction confirmed that the cryogel material was amorphous. In addition, the column presented a hydraulic permeability of 4.15 × 10−13 m2, axial dispersion ranging from 10−7 to 10−6 m2 s−1, and a height equivalent to a theoretical plate ranging from 0.10 to 0.21 cm. The highest adsorption of lysozyme (67.65 mg g−1) was obtained using sodium thiocyanate saline solution (0.025 mol L−1, pH 5.0). The ability of the cryogel‐Phe column to capture and purify lysozyme was confirmed by high enzymatic activity (1294.17 U ml−1), purity (87.92%), purification factor (11.49), and sulphate‐polyacrylamide electrophoresis gel (SDS‐PAGE) electrophoresis gel.

Integrated in silico and experimental discovery of trimeric peptide ligands targeting Butyrylcholinesterase

Butyrylcholinesterase (BChE) is recognized as a high value biotherapeutic in the treatment of Alzheimer’s disease and drug addiction. This study presents the rational design and screening of an in-silico library of trimeric peptides against BChE and the experimental characterization of peptide ligands for purification. The selected peptides consistently afforded high BChE recovery (> 90 %) and purity, yielding up to a 1000-fold purification factor. This study revealed a marked anti-correlated conformational movement governed by the ionic strength and pH of the aqueous environment, which ultimately controls BChE binding and release during chromatographic purification; and highlighted the role of residues within and allosteric to the catalytic triad of BChE in determining biorecognition, thus providing useful guidance for ligand design and affinity maturation.

Reactive Green 19 dye-ligand immobilized on the aminated nanofiber membranes for efficient adsorption of lysozyme: Process development and optimization in batch and flow systems

The performance of lysozyme adsorption by the aminated nanofiber membrane immobilized with Reactive Green 19 (RG19) dyes was evaluated in batch and flow systems. The physicochemical properties of the dye-immobilized nanofiber membrane were characterized. The parameters of batch-mode adsorption of lysozyme (e.g., pH, initial dye concentration, and lysozyme concentration) were optimized using the Taguchi method. In a flow process, the factors influencing the dynamic binding performance for lysozyme adsorption in the chicken egg white (CEW) solution include immobilized dye concentration, adsorption pH value, feed flow rate, and feed CEW concentration. The impact of these operating conditions on the lysozyme purification process was investigated. Under optimal conditions, the recovery yield and purification factor of lysozyme achieved from the one-step adsorption process were 98.52% and 143 folds, respectively. The dye-affinity nanofiber membrane also did not exhibit any significant loss in its binding capacity and purification performance after five consecutive uses.

Recovery of acid proteases from fishery discards with aqueous micellar two-phase systems and their use for X-ray film recycling

The continuous expansion of fish production results in increasing amounts of discards, i.e. viscera, skin and frame, whose inappropriate disposal causes environmental damage. Conversion of this waste into value-added products is one of the best alternatives to reduce the pollution problem. The feasibility of using aqueous micellar two-phase systems (AMTPS), formed by biodegradable Genapol X-080 (GX-080) and sodium citrate, to recover valued acid proteases from surubí (Pseudoplatystoma sp) fishery discards was evaluated for the first time. The effects of biomass load, pH and surfactant concentration on the extraction were analyzed using a two-level full factorial design. GX-080 concentration and biomass load were the most significant factors (p < 0.05). AMTPS formed by GX-080 4% w/w showed a successful performance for the recovering of 69% of AP from the crude extract in the surfactant-rich phase with a purification factor of 1.83. Regard to the storage stability, this extract conserved at least 89% of its proteolytic activity for 76 days at 0 °C. It also showed a notable hydrolyzing activity of the gelatin-silver coating of used X-ray films, thus opening new perspectives on the use of acid proteases for recycling purposes.

Cloud point extraction based on non-ionic surfactants: An ecofriendly tool for recovering papain from papaya latex

The potential use of cloud point extraction with aqueous micellar two-phase systems (AMTPS), formed by the non-ionic biodegradable surfactants Triton X-114 (TX-114) and Genapol X-080 (GX-080), to recover papain (PAP) from fresh papaya latex was evaluated. Binodal curves obtained for both surfactants, in absence and presence of PAP, showed cloud points below 40 °C, the PAP presence slightly favoring the phase separation phenomenon. Partitioning experiments showed that PAP migrated toward the micelle-poor phase independently of the surfactant and the working temperature assayed, this behavior being consistent with this enzyme hydrophilic character. PAP recoveries of over 60% in the poor-micelle phase were obtained when partitioned fresh papaya latex in both surfactant-based AMTPS. TX-114 systems (4.00% w/w, 33 °C) allowed recovering 66% of PAP with a purification factor of 4.6. On GX-080 systems (4.00% w/w, 42 °C) a yield of 78% PAP and a purification factor of 3.7 were achieved. This AMTPS also allowed the separation of PAP from reducing sugars present in papaya latex. These results show that TX-114 and GX-080-based AMTPS could be potentially used as simple and economical tools at recovering PAP from papaya latex.

Recovery of nisin from culture supernatants of Lactococcus lactis by ultrafiltration: Flux properties and separation efficiency

Nisin is a class I bacteriocin, which is produced by lactic acid bacteria (LAB) belonging to Lactococcus and Streptococcus genera. Nisin is approved by the FDA (Food and Drug Administration) as a natural biopreservative agent under reference E234. Besides food application, nisin has also a potential for therapeutic application to treat a range of infectious diseases. Nisin, which is commercially available, is mainly produced by liquid fermentation. The production of commercial high-quality nisin is hampered by the high cost of downstream processing, which often involves subsequent steps of salt precipitation, centrifugation, and chromatography. Membrane processes offer a set of advantages. Indeed, these processes are simple and effective technologies for protein concentration and separation; One of the biggest limitations is the decrease in permeate flux. In this study, the performance of three UF membranes (10, 50, and 100 kDa) for the separation and concentration of nisin obtained from Lactococcus lactis (L. lactis) culture supernatants was evaluated. Permeate flux decline, fouling resistances, fouling index, nisin recovery in the permeate and retentate streams were evaluated. The 10 kDa membrane showed the best nisin recovery properties with the highest recovery yield and purification factor, achieving 100% and 4 in the retentate stream, respectively.

Non-Cryoprecipitation Separation of Coagulation FVIII and Prothrombin Complex Proteins by Pseudoaffinity Calcium Elution Chromatography Using Anion Exchange Resin.

Hemophilia A is treated with human plasma coagulation factor VIII (FVIII) replacement therapy and Hemophilia B with coagulation factor IX, which is purified from prothrombin complex concentrate (PCC). In this paper we evaluated the separation of FVIII and PCC by directly loading raw thawed plasma to an anion exchange resin (AEX). Under this relatively high ionic strength, most of the plasma proteins such as albumin, immunoglobulins and others were not adsorbed. Five resins commonly used in protein purification (plasma fractionation) were tested. With all resins, PCC was eluted by pseudoaffinity in a calcium gradient step. Afterwards, FVIII could be recovered with a good yield and high purification factor in the salt gradient step with 400–500 mM NaCl. Using ANX Sepharose FF and Q Sepharose FF, the CaCl2 elution step was introduced after the intermediate wash with 200 mM NaCl, whereas using DEAE Sepharose FF, Fractogel EMD TMAE and Fractogel EMD DEAD, PCC eluted after the wash of the unbound proteins. Our results indicate that three important fractions: (1) albumin, immunoglobulin etc.; (2) PCC; and (3) FVIII can be separated in one chromatographic AEX column and the delicate and troublesome cryoprecipitation can be eliminated, making the purification of blood products faster and cheaper.

Characterization and optimization of polymer-polymer aqueous two-phase systems for the isolation and purification of CaCo2 cell-derived exosomes.

Exosomes are cell-derived vesicles that present attractive characteristics such as nano size and unique structure for their use as drug delivery systems for drug therapy, biomarkers for prognostic, diagnostic and personalized treatments. So far, one of the major challenges for therapeutic applications of exosomes is the development of optimized isolation methods. In this context, aqueous two-phase systems (ATPS) have been used as an alternative method to isolate biological molecules and particles with promising expectations for exosomes. In this work, fractionation of exosomes obtained from CaCo2 cell line and culture media contaminants were individually performed in 20 polymer-polymer ATPS. The effect of design parameters such as polymer composition, molecular weight, and tie-line length (TLL) on polyethylene glycol (PEG)-Dextran, Dextran-Ficoll and PEG-Ficoll systems was studied. After partition analysis, 4 of the 20 systems presented the best exosome fractionation from contaminants under initial conditions, which were optimized via salt addition (NaCl) to a final concentration of 25 mM, to improve collection efficiency. The PEG 10,000 gmol-1 –Dextran 10,000 gmol-1 system at TLL 25% w/w with NaCl, showed the best potential isolation efficiency. Following this proposed strategy, an exosome purification factor of 2 in the top PEG-rich phase can be expected furtherly demonstrating that ATPS have the potential for the selective recovery of these promising nanovesicles.

Freeze-thaw-assisted aqueous two-phase system as a green and low-cost option for analytical grade B-phycoerythrin production from unicellular microalgae Porphyridium purpureum

Microalgae are promising alternative sources for natural phycobiliprotein production since they grow rapidly and can be cultured on a large scale. However, the overall economic viability of analytical grade phycoerythrin production from microalgae is limited by the high cost of the pretreatment required to break down the cell wall, as well as the expensive reagents and equipment used during purification. This study developed a novel extraction and purification process to recovery B-phycoerythrin (B-PE) in a freeze-thaw-assisted aqueous two-phase system (FT-ATPS) from Porphyridium purpureum. The effects of culture strategy, extraction method and purification factors on B-PE production were studied systematically. The results revealed that low light induced B-PE synthesis, whereas hindered the growth. The stepwise light supplement strategy alleviated the growth limitation under low-light conditions and increased the final B-PE production, up to 182.1 ± 4.1 mg/L (increasing by 73.1 %). Compared with the traditional electricity highly-consumed ultrasonication method, the green freeze-thaw method could extract B-PE without damaging the cell structure and save much more power (>98 %), and the extraction rate reached the highest when frozen at −80 °C. The purity and concentration of B-PE in crude extract were proved to have a significant effect on ATPS purification, and the higher the former, the better the latter. A total production cost of 0.045 $/mg (decreasing by 96.2 %) and purity of 5.6 ± 0.2 (A545/A280) for analytical grade B-PE production were achieved by FT-ATPS based on 4 % polyethylene glycol (PEG2,000) and 20 % ammonium sulfate. Relative to the conventional method of phycoerythrin production, the method developed herein has greater safety, lower cost, and more environmental benefits, which can be function appropriately in the expensive purification of analytical grade phycobiliproteins.

One-step salting-out extraction of bacteriophage from its infection broth of Acinetobacter baumannii

Phages as a potential alternative antibiotic for multidrug-resistant bacterial infections are receiving great attention worldwide. However, the traditional separation and purification of phage are cumbersome, time-consuming, costly and inefficient. In this study, phage phiAB9 for multidrug-resistant Acinetobacter baumannii was separated and purified by a simple and cost-saving one-step salting-out extraction (SOE). Several kinds of salts and organic solvents without effect on phage survival were chosen to form the SOE systems, and the composition of SOE systems were optimized according to the phage recovery rate and impurity removal rate. After one-step SOE by an optimal system composed of 18% (w/w) ammonium citrate and 40% (w/w) ethyl acetate, the recovery rate of phage in the middle phase could reach to 90.82%, and most proteins (99.57%), cells (97.98%) and endotoxin (84.08%) were well removed, with a concentration factor of 210 and the purification factors of phage to proteins, cells and endotoxin were 303.64, 133.55 and 5.36, respectively. Comparing with two-step SOE and traditional aqueous two-phase extraction, one-step SOE may be an available method for the separation and purification of phages.

Adsorption and immobilization of β-glucosidase from Thermoascus aurantiacus on macroporous cryogel by hydrophobic interaction

Enzyme immobilization has been reported as a promising approach to improving parameters such as thermal stability, pH and reusability. In this study, a polyacrylamide cryogel functionalized with L-phenylalanine was prepared to be used in the adsorption of β-glucosidase from Thermoascus aurantiacus, aiming at its separation and also its immobilization on the cryogel matrix. The enzyme was produced by solid state fermentation. First, the adsorption was studied as a function of the pH and the resulting yield (Y, %) and purification factor (PF , dimensionless) were determined (1.57–5.13 and 64.19–91.20, respectively). The PF and yield from eluate samples obtained at pH 3.0 were the highest (5.13 and 91.20, respectively). Then, β-glucosidase was immobilized on the hydrophobic cryogel and the recovery activities (%) were determined as a function of temperature and in the presence of different saline solutions. The values ranged from 14.45 to 45.97. As expected, salt type and ionic strength affected the activity remained in the immobilized β-glucosidase. The average bioreactor activity was 39.9 U/g of dry cryogel and its operational stability was measured, with no decrease in activity being observed during seven cycles. Kinetic parameters of free and immobilized enzyme were determined according to different models.

Downstream Process of Xylanase Production from Oil Palm Empty Fruit Bunches: A Review

As an enzyme that can hydrolyze the glycosidic bond of hemicellulose, xylanase is widely used in pulp and paper, feed, food, and beverage industries. The vast availability of lignocellulosic biomass waste offers the potential of xylanase production by microorganisms via submerged and solid-state fermentation. However, the specific activity of the produced crude enzyme is still too low to be directly used in a commercial application. Downstream processing of the enzyme, that is purification and concentration, is necessary. Centrifugation or filtration can be used as a pre-treatment process for biomass separation before the next purification technologies. Precipitation, aqueous two-phase system (ATPS), chromatography, and membrane ultrafiltration (UF) are technologies for enzyme purification. Freeze drying can be used also as a late-stage purification procedure. ATPS and membrane UF have been emerged as a recovery method enabling the purification and concentration of the target biomolecule in a single operation. The downstream process of an enzyme usually accounts for 70-90% of the total production cost. Nonetheless set of purification methods that have high purification factors, cost-effective, high productivity, and environmentally benign strategies are needed such that the xylanase production process can be conducted economically at an industrial/commercial scale. This paper reviews processes for xylanase purification/concentration produced by using solid-state fermentation configuration. A membrane-based process for downstream processing of xylanase production is suggested.

Sea-Urchin carbon nitride with carbon vacancies (C-v) and oxygen substitution (O-s) for photodegradation of Tetracycline: Performance, mechanism insight and pathways

The fast recombination of electron-hole pairs results in low photoactivity, limiting carbon nitride further application in wastewater purification. To address this issue, regulation of electron configuration via optimizing surface atom properties is a promising strategy for improving the separation of electron-hole pairs. Here, sea-urchin carbon nitride (SUCN-6) with C-v and O-s mediated electronic structures were facile prepared, endowing it with a decrease in transportation resistance of charge carriers and increased activation towards oxygen. Under visible light, the SUCN-6 showed a removal efficiency of ∼ 89% towards tetracycline (TC) in 20 min and no obvious decaying after five consecutive cycles. This TC photodegradation follows a first-order kinetic, and its kinetic constant reaches 0.0908 min−1, which is 2.35 folds higher than that of pristine carbon nitride. In addition, several crucial factors of practical wastewater purification (such as pH, TC concentration, water sources, inorganic salts, and organic matters) that might affect photoactivity were investigated. Moreover, the degradation pathways of TC and intermediates were detected by an HPLC-MS and electron spin resonance instrument, and a possible photodegradation mechanism over SUCN-6 was elucidated. This study offers new insights into the photocatalysis kinetics and photodegradation mechanism over carbon nitride for environmental remediation.