Rapeseed Protein Research Articles

Enzyme‐catalyzed acylation improves gel properties of rapeseed protein isolate

Although rapeseed protein isolate (RPI) possessed some good functional properties, the use of RPI as an ingredient in the food industry is restricted mainly due to its inferior gelation. The purpose of this study was to improve the heat-induced gel properties of RPI using double processes of acylation and additional transglutaminase catalysis. Scanning electron microscopy showed that the gel formed by native RPI exhibited randomly aggregated particulate network structures whereas transglutaminase (TG)-assisted RPI gels significantly improved gelation properties. More importantly, the combined modifications of RPI using TG-assisted acylation can form a gel with unique percolating and small porous structure. Furthermore, TG-catalyzed 5% acylated RPI gel (100 U g-1 , protein basis) exhibited excellent gel properties in terms of gel strength, thermal stability, surface roughness and apparent viscosity compared to non-treated or single modification of RPI gel as determined by texture analyzer, atomic force microscopy and rheometer. Mechanistically, Fourier-transform infrared spectra and gel dissociation test revealed that TG-catalyzed acylation extensively unfolded the hydrophobic and sulfhydryl residues of RPI, in turn, reinforced re-assembly of protein molecules via hydrophobic interactions and disulfide bonds during gel formation. Combined processes of acylation and additional TG catalysis improved the thermal gelation properties by altering inter- and intra-protein structures. Such sequential processes will provide a promising approach to improve the protein gelation that could be potentially applied in the food industry. © 2020 Society of Chemical Industry.

Effect of pH regulation on the components and functional properties of proteins isolated from cold-pressed rapeseed meal through alkaline extraction and acid precipitation

Cold-pressed rapeseed meal with high protein content (38.76% protein dry weight basis) was used to prepare rapeseed protein isolates (RPIs) by alkaline extraction (pH 8.0, 9.0, 10.0, 11.0, 12.0 and 13.0) and acid precipitation (pH 3.0, 3.5, 4.0, 4.5, 5.0 and 5.5). The protein with an intact structure and the highest yield (65.08%) was obtained at extraction pH 9.0 and precipitation pH 4.5, accompanied by the lowest D-amino acid content, the lightest colour and the lowest contents of glucosinolates (2.85 mmol/kg), phytic acid (1.05 mg/g) and sinapine (0.68 mg/g). Additionally, water/oil absorption, foaming and emulsifying capacities decreased with decreasing precipitation pH, while the solubility showed the reverse trend. During gastric simulation digestion, the α-polypeptide of cruciferin and napin in the RPIs showed digestive resistance. Overall, pH regulation might be an effective method to isolate high quality RPIs for use in the food processing industry.

Effect of Acylation of Rapeseed Proteins with Lauroyl and Oleoyl Chloride on Solubility and Film-Forming Properties

Rapeseed proteins show good film-forming properties, giving them a promising potential as bio-based ingredients for the technical industry, e. g. for films and coatings. However, their hydrophilicity often poses problems in regard to water-stability of coatings. Protein modification using fatty acids is known to reduce hydrophilicity, however, it has not been tested to improve film-forming properties of rapeseed proteins before. In the present study, a rapeseed protein concentrate (RPC) was acylated at low and high modification degree using lauroyl chloride and oleyol chloride. The protein solubility was determined and the modified RPCs were used for the preparation of cast-films to measure the changes of mechanical properties (tensile strength, elongation at break), surface energy, oxygen permeability and light transmission. The protein solubility in water was lowered from 100% for the non-modified RPC to < 15% for highly modified RPCs at pH 7. The tensile strength of films increased by factors of 3.5 and 4 for highly modified samples, respectively. Surface energy and oxygen permeability revealed an increase of hydrophobicity that correlated with the modification degree. The light transmission was reduced by modification. The results confirm the increased hydrophobicity of acylated RPCs and demonstrate the potential of modified rapeseed proteins as an ingredient for technical products, such as packaging layers, coatings and adhesives.Graphic

Rapeseed Protein Nanogels As Novel Pickering Stabilizers for Oil-in-Water Emulsions.

Recently plant protein Pickering particles have received tremendous interests because of their environmentally friendly, biodegradable, and safe characteristics. However, developing plant protein particles as stabilizers of Pickering emulsion still face many challenges. In current study, a novel nanogel system produced from acylated rapeseed protein isolates (ARPI) was used to stabilize Pickering emulsions. Results showed that self-assembled nanogel after native RPI modified by acylation adjusted the three-phase contact angle of ARPI nanogels system to 86.7° closing to a neutral wettability. At constant oil phase fraction (0.3, v/v), increasing the ARPI nanogels concentrations produced smaller droplet sizes of Pickering emulsions, whereas all freshly prepared Pickering emulsions were stable except 0.1% (w/v) ARPI nanogel-stabilized Pickering emulsion occurred with creaming. The rise of the oil phase fraction showed little influences on the droplets size and visual appearances of Pickering emulsions at a fixed ARPI nanogels concentration (0.75%, w/v). Moreover, the prepared ARPI nanogels stabilized Pickering emulsions were stable against aggregations of droplets at a range of pH conditions ranging from 5.5 to 8.5 and salt concentration as high as 0.2 M. Additionally, the ARPI nanogels concentration above 0.5% favored the formation of Pickering emulsion with long-term storage stability (up to 30 days) against creaming. Microscopic images evidenced that ARPI nanogels could absorb and anchor at the droplets surface forming an interfacial layer. Above findings may deliver a potential strategy for fabricating stable Pickering emulsion based on plant protein particles and are of important significance for the utilization of rapeseed protein in the food industry.

The Effects of Oil Extraction Methods on Recovery Yield and Emulsifying Properties of Proteins from Rapeseed Meal and Press Cake.

The agricultural sector is thought to be responsible for around 30% of the anthropogenic climate change and it is well established that high meat consumption has a tremendous impact on the environment. Rapeseed is mainly used for production of vegetable oil, but press cake has high protein content with the potential for incorporation into new plant protein-based foods. Protein was recovered from press cakes generated from different oil pressing processes. Industrially cold-pressed, hot-pressed, and solvent-extracted rapeseed press cake and the effect of heat treatment in the recovery process was assessed. Protein recovery yield, protein concentration and emulsifying properties were analyzed. Cold-pressed rapeseed press cake (RPC) recovered in the absence of heat, yielded the highest protein recovery (45%) followed by hot-pressed rapeseed meal (RM) (26%) and solvent-extracted RM (5%). Exposure to heat during recovery significantly reduced the yield for cold-pressed RPC but no difference was found for hot-pressed RM. The protein recovery yield was improved for solvent-extracted RM when heat was applied in the recovery process. The ability to stabilize emulsions was highest for protein recovered from cold-pressed RPC, followed by hot-pressed RM and solvent-extracted RM, and was in the same range as commercial emulsifying agents. Heat treatment during recovery significantly reduced the emulsifying properties for all pressing methods examined. This study suggests that cold-pressed rapeseed press cake without heat in the recovery process could be a successful strategy for an efficient recovery of rapeseed protein with good emulsifying properties.

Protein Recovery from Rapeseed Press Cake: Varietal and Processing Condition Effects on Yield, Emulsifying Capacity and Antioxidant Activity of the Protein Rich Extract.

Protein was recovered from five varieties and a mixed blend of cold-pressed rapeseed press cake by leaching and precipitation in a water-based process, and the protein recovery yield varied from 26–41% depending on variety. Exposure for heat during protein recovery severely reduced the rapeseed proteins’ ability to stabilize the oil–water interface of emulsion droplets. Protein extract from Lyside had the best emulsifying properties of the varieties investigated. Oxidation rate was assessed by the Thiobarbituric Acid Reactive Substances (TBARS) method and rapeseed protein extracts from Epure and Festivo had higher capacity to delay oxidation compared with soy lecithin. There are possibilities to broaden the use of rapeseed whereby recovered rapeseed protein can be used as a plant-based multifunctional ingredient with emulsifying capacity and which has a delaying effect on oxidation.

Effects of dietary rapeseed glucosinolates, sinapic acid and phytic acid on feed intake, growth performance and fish health in turbot (Psetta maxima L.)

Various antinutritional factors (ANF) present in rapeseed protein products can adversely affect the voluntary feed intake, growth performance and physiology as well as health of fish. The aim of the presented study was to evaluate which ANF is primarily responsible for the mentioned effects. Six identical semi-purified diets based on casein were produced. One unsupplemented basal diet (BD) served as control. The remaining five diets were supplemented with graded levels of glucosinolates (low = GL, high = GH), sinapic acid (low = SL, high = SH) and one level of phytic acid (PA). Diets were fed to triplicate groups of juvenile turbot (Psetta maxima L.) until apparent satiation for 50 days. Voluntary feed intake, growth performance as well as physiology of fish were investigated. Feed intake and growth performance of fish did not differ significantly (p < 0.05) between experimental groups. However, the feed conversion ratio (FCR) increased significantly due to dietary phytic acid supplementation (10 g kg−1feed). Also the protein efficiency ratio (PER) showed a strong tendency to decrease in fish fed diet PA (p = 0.0518). Furthermore no significant differences were detected among dietary treatments regarding the proximate final body composition, hepatosomatic index, splenic index and haematocrit of fish. Concentrations of the thyroidal hormones thyroxine (T4) and triiodothyronine (T3) in blood plasma were unaffected in all fish fed experimental diets. It was concluded that higher dietary levels of phytic acid, similar to commercial plant-based diets, can adversely affect nutrient utilisation in fish nutrition. Other investigated ANF did not harm fish performance significantly in applied concentrations when supplemented in a pure grade.

Development of food products with addition of rapeseed presscake fermented by Rhizopus

Purpose The purpose of this paper is to explore the prospects and limitations of consumer acceptance towards food products that have been enriched with rapeseed protein in the form of fermented rapeseed presscake. Design/methodology/approach In total, 16 different food products with up to 18 per cent fermented rapeseed presscake were developed. Three types of vegetarian spreads and one type of bread were formulated, and consumer acceptance was assessed by sensory evaluation, questionnaires and in a focus group. Findings The fermentation of the rapeseed presscake by using the tempeh mould, Rhizopus oligosporus, had little if any effect on the bitterness of the presscake, and the bitter taste and aftertaste of the fermented presscake limited its use in food products. However, promising results were obtained when the breads and vegetarian spreads that were prepared with 5–6 per cent dried fermented rapeseed presscake were presented to sensory panels and to consumers, provided that the bitterness was masked, to some degree, by appropriate formulations and processes. Research limitations/implications Responders consisted mainly of younger people open to sustainable, plant-based nutrition, and may not represent the general population. Nevertheless, the results have implications for further research on utilization of rapeseed proteins. Practical implications It is possible to use the fermented rapeseed presscake to enrich various foods with protein or to replace other proteins, e.g. from animals or soybeans, provided that the degree of degradation of undesired compounds can be better standardized. Originality/value This study provides useful information for how to increase the use of underutilized plant proteins for human nutrition.

Effects of Succinylation on the Physicochemical Properties and Structural Characteristics of Edible Rapeseed Protein Isolate Films

AbstractThis study described the manufacture and characterization of different levels of succinic anhydride (0–15%, w/w) on rapeseed protein isolate (RPI) films. Mechanical, barrier, optical, surface, microstructure, and thermal properties were investigated. Results showed that 5% succinylated rapeseed protein isolate (SRPI) film revealed the overall optimal properties. The mechanical and barrier properties of 5% SRPI film were enhanced significantly compared to the RPI film. The water vapor permeability values of RPI and 5% SRPI films were 0.77 and 0.32 g mm/[hour m2 kPa], respectively. Succinylation improved the color of RPI. The contact angle of 5% SRPI film was 108.68° and was the most hydrophobic combination. Moreover, scanning electron microscopy images revealed that 5% SRPI showed a dense, homogenous, and smooth microstructure. Thermogravimetric analysis results verified a low level of succinylation strengthened the thermal resistance of SRPI films. The content of β‐sheet was almost half of all secondary structures after succinylation, which was supported by analyses of Fourier transform infrared spectroscopy. Results demonstrated that succinylation could be employed to improve the properties of rapeseed protein and SRPI films have great potential to be used as edible films.

Improved in vitro digestibility of rapeseed napin proteins in mixtures with bovine beta-lactoglobulin.

Mixing of different protein sources can lead to either predictable, synergistic, or antagonistic effects on the protein digestibility. This study investigated the in vitro protein digestibility (IVPD) of protein mixtures between a napin-rich rapeseed (Brassica napus L.) protein concentrate (RP2) and bovine milk whey proteins (WPs; α-LA, alpha-lactalbumin; β-LG, beta-lactoglobulin) at mixing ratios of 20:80, 40:60, 60:40, and 80:20 w/w protein. Enzymatic hydrolysis consisted of pepsin digestion (1 h) followed by short- (+1 h), medium- (+3 h), or long-term (+24 h) pancreatin digestion. IVPD was differentially affected by the WPs type, mixing ratios, and total hydrolysis times. RP2/β-LG protein mixtures showed a partially synergistic effect at mixing ratios of 40:60 and 60:40 w/w, leading to an increased short-term IVPD of 7-10%. LC-MS analysis revealed a markedly improved short-term digestibility of the napin proteins when combined with bovine β-LG. This study demonstrated that specific mixtures between animal and plant protein sources exhibit an improved digestibility due to synergistic protein-protein interactions.

RAP-8 ameliorates liver fibrosis by modulating cell cycle and oxidative stress

AimsThe rapeseed protein derived peptide DHNNPQIR (named as RAP-8) has been previously reported to possess antioxidant activity and alleviate liver fibrosis. The purpose of the present study was to investigate the potential crucial pathways involved in ameliorating liver fibrosis of RAP-8. Main methodsNext-generation sequencing of messenger RNA (RNA-Seq) analysis of the fibrotic and RAP-8 treated mice was performed. Western blot, qPCR and flow cytometry detection analysis were conducted to measure cell cycle and oxidative stress in LX-2 cells and liver samples. Key findings588 overlapped differentially expressed genes were obtained from a batch of genes RAP-8 altered. Gene Ontology enrichment analysis revealed that changes in the most significant modules were mainly enriched in cell division, nuclear division and mitotic cell cycle process, while alterations in Kyoto Encyclopedia of Genes and Genomes were mainly enriched in cell cycle. Thereafter, according to the co-expression network analysis, the regulations of three core genes (Cenpp, Cyp2c55, Serpinh1) were verified that might be targets for treating liver fibrosis. Furthermore, through experimental verification, we demonstrated that RAP-8 induced cell cycle arrest and prevents oxidation stress. SignificanceAs a promising therapeutic candidate for hepatic fibrosis treating, RAP-8 exhibited anti-fibrotic effects via exerting cell cycle arrest and inhibiting oxidative stress.

Rapeseed (Brassica napus L.) biofortification with selenium: How do sulphate and phosphate influence the efficiency of selenate application into soil?

ABSTRACTThe connection between sulphur (S) and selenium (Se) metabolism makes rapeseed (Brassica napus) an attractive candidate for Se fortification. Unfortunately, fertilizers may interfere with the availability of selenate (SeO42-) in numerous ways, including both soil and physiological processes. Experiments on two agricultural soils amended with SeO42- (32 μg Se kg−1 soil), sulphate and phosphate (each at three levels of supply) were established to elucidate the effect of these anions on the selenization efficiency. Maximal efficiency in Chernozem soil was roughly two-fold higher (455 μg Se kg−1 seed) than in Cambisol. Sulphate significantly decreased (up to 28%) the seed Se contents in Cambisol, while an enhancement (up to 33%) was found in Chernozem. In the Chernozem, the induction of collective S and Se translocation toward the seed more than compensated for any competition effects due to the highest sulphate supply. In Cambisol, plant Se distribution did not follow that of S as closely as in Chernozem. Phosphate did not significantly alter the fortification efficiency. Resistance of rapeseed proteins to protease hampered a quantitative investigation of changes in Se speciation under different S supplies. Nevertheless, protein-bound selenomethionine was the predominant Se storage form and traces of other Se species were also identified.

Rapeseed protein concentrates for non-food applications prepared from pre-pressed and cold-pressed press cake via acidic precipitation and ultrafiltration

Rapeseed protein concentrates (RPCs), prepared from residual press-cakes, show excellent techno-functional properties, such as emulsification and foaming. However, food application is hindered by residual contents of anti-nutritive components. Limited research has been invested in studying RPCs as a potential ingredient in non-food applications. The present study reports RPC preparation from cold-pressed rapeseed meal (CPM) and pre-pressed rapeseed meal (PPM) as a potential ingredient for the chemical industry. Protein isolation was achieved after extraction at pH 5.7–7.0 via i) ultrafiltration or ii) a combination of acidic precipitation followed by ultrafiltration. Protein isolation yields obtained from CPM (36.5%–40.6%) were 60%–90% higher compared to PPM (19.4%–26.0%), highlighting the positive effect of low-temperature processing during defatting. Protein contents of RPCs were 75.3%–87.1% and were highest for preparations obtained through precipitation. RPCs obtained through ultrafiltration showed good solubility properties and very high values for emulsifying capacity (688–768 mL/g) as well as foaming activity (1834%–2834%). In contrast, precipitated RPCs showed lower functional values (410–445 mL/g, 888%–938%, respectively). All RPCs had excellent film-forming properties in cast-film experiments. Thus, RPCs are promising ingredients for industrial non-food applications such as adhesives, detergents, paints, varnishes and biodegradable polymers.

Hydrolysis of Soybean and Rapeseed Proteins with Enzyme Complex Extracted from the Pyloric Caeca of the Cod

Hydrolysis of Soybean and Rapeseed Proteins with Enzyme Complex Extracted from the Pyloric Caeca of the Cod

Digestion dynamics in broilers fed rapeseed meal

Rapeseed proteins are described to be poorly digestible in chickens. To further identify some molecular locks that may limit their use in poultry nutrition, we conducted a proteomic study on the various chicken digestive contents and proposed an integrative view of the proteins recruited in the crop, proventriculus/gizzard, duodenum, jejunum, and ileum for digestion of rapeseed by-products. Twenty-seven distinct rapeseed proteins were identified in the hydrosoluble fraction of the feed prior ingestion. The number of rapeseed proteins identified in digestive contents decreases throughout the digestion process while some are progressively solubilized in the most distal digestive segment, likely due to a combined effect of pH and activity of specific hydrolytic enzymes. Fifteen chicken proteins were identified in the hydrosoluble proventriculus/gizzard content, including chymotrypsin-like elastase and pepsin. Interestingly, on the 69 distinct proteins identified in duodenum, only 9 were proteolytic enzymes, whereas the others were associated with homeostasis, and carbohydrate, lipid, vitamin and hormone metabolisms. In contrast, chicken proteins identified in jejunal and ileal contents were mostly proteases and peptidases. The present work highlights the relevance of using integrative proteomics applied to the entire digestive tract to better appreciate the protein profile and functions of each digestive segment.

Rapeseed protein-derived peptides, LY, RALP, and GHS, modulates key enzymes and intermediate products of renin\u2013angiotensin system pathway in spontaneously hypertensive rat

Rapeseed proteins are a rich source of bioactive peptides. LY, RALP and GHS were previously identified from rapeseed protein hydrolysates as potent ACE and renin inhibiting peptides. In this study, the rapeseed peptides were individually evaluated for their molecular mechanisms and regulatory effects on components of the renin–angiotensin system in spontaneously hypertensive rats (SHR), including the mRNA and/or protein levels of angiotensin-converting enzyme (ACE), renin, ACE2, angiotensin II and angiotensin-(1–7) in myocardial tissues. Oral administration of 30 mg peptides/kg body weight every 2 days for five weeks significantly decreased the systolic blood pressure and the myocardial mRNA and protein levels of ACE and renin in SHR. LY, RALP and GHS also increased the expression of ACE2, angiotensin-(1-7) and Mas receptor levels, which may have mediated their antihypertensive activity. Dipeptide LY also inhibited angiotensin II protein expression in the heart tissue. Taken together, the finding demonstrates the multi-target physiological effects of the rapeseed peptides, beyond ACE and renin inhibition, which enhances knowledge of the antihypertensive mechanisms of food protein-derived peptides.

Rapeseed Protein-Derived Antioxidant Peptide RAP Ameliorates Nonalcoholic Steatohepatitis and Related Metabolic Disorders in Mice.

Rapeseed protein hydrolysates have recently shown in vitro antioxidant and anti-inflammatory activities. However, scant data exist about their in vivo activities. Here, we report that the peptide DHNNPQIR (hereinafter referred to as RAP-8), a bioactive peptide originated from rapeseed protein, exhibits excellent in vivo efficacy in mouse models of nonalcoholic steatohepatitis (NASH) and hepatic fibrosis. We demonstrated that RAP-8 significantly reduced hepatic steatosis and improved insulin resistance and lipid metabolism. Furthermore, RAP-8 showed markedly reduced hepatic inflammation, fibrosis, liver injury, and metabolic deterioration. In particular, RAP-8 directly suppressed fibrosis-associated gene expression, including α-smooth muscle actin (α-Sma) and collagen type I (Col-1α) in the liver of mice in vivo. In addtion, RAP-8 significantly decreased macrophage infiltration and reduced pro-inflammatory cytokines secretion. Finally, we found that RAP-8 administration significantly decreased oxidative stress-induced apoptosis in liver injury induced by CCl4. Therefore, our results suggest that RAP-8 could be available for treatment of NASH and NASH-related metabolic disorders as a potential therapeutic candidate.

Kaempferol 3- O-(2‴- O-Sinapoyl-β-sophoroside) Causes the Undesired Bitter Taste of Canola/Rapeseed Protein Isolates.

By means of activity-guided fractionation using taste dilution analysis, liquid chromatography-tandem mass spectrometry (LC-MS/MS), liquid chromatography-time-of-flight mass spectrometry, one-/two-dimensional nuclear magnetic resonance spectroscopy, LC-MS/MS quantitation, dose-over-threshold considerations, and sensory spiking experiments, kaempferol 3- O-(2‴- O-sinapoyl-β-sophoroside), exhibiting a bitter taste above the low threshold concentration of 3.4 μmol/L, was found for the first time as the key molecule contributing to the unpleasant bitter taste of rapeseed (canola) protein isolates. This finding opens new avenues for a biorefinery approach targeting an off-taste removal.

Controlled fermentation of rapeseed presscake by Rhizopus, and its effect on some components with relevance to human nutrition

The use of rapeseed protein could contribute to meeting the increasing demand for plant proteins with high biological value in human nutrition. In order to make rapeseed presscake fit for human consumption, the presscake was fermented by using the tempeh mould, Rhizopus microsporus var. oligosporus. Fermentation was satisfactory at initial levels of added acetic acid of 40–60 mmoles/Kg, aw of 0.97, pasteurization, surface inoculation and incubation at 32 °C and 90–95% relative humidity. It was crucial to stop the fermentation once the mould had grown and metabolized sufficiently but before a major rise in pH and subsequent growth of acid-sensitive sporeforming bacteria occurred. Some degradation of glucosinolates, cell wall polysaccharides and phenolic compounds was found, but there was some evidence that growth and metabolism of the mould also depended on the texture of the presscake, as these factors affect the oxygen supply to the mould. In conclusion, it is possible to ferment rapeseed presscake by using the “Tempeh starter” Rhizopus oligosporus, but in order to use the resulting product to enrich various foods with protein or replace other proteins, the degree of degradation of undesired compounds should be further standardized, especially by the control of the pH, oxygen supply, and fermentation time.

In Situ Proapoptotic Peptide-Generating Rapeseed Protein-Based Nanocomplexes Synergize Chemotherapy for Cathepsin-B Overexpressing Breast Cancer.

Intracellular activation of nanomaterials within cancer cells presents a powerful means to enhance anticancer specificity and efficacy. In light of upregulated lysosomal protease cathepsin-B (CathB) in many types of invasive cancer cells, herein, we exploit CathB-catalyzed biodegradation of acetylated rapeseed protein isolate (ARPI) to design polymer-drug nanocomplexes that can produce proapoptotic peptides in situ and synergize chemotherapy. ARPI forms nanocomplexes with chitosan (CS) and anticancer drug doxorubicin (DOX) [DOX-ARPI/CSnanoparticles (NPs)]by ionic self-assembly. The dual acidic pH- and CathB-responsive properties of the nanocomplexes and CathB-catalyzed biodegradation of ARPI enable efficient lysosomal escape and nuclei trafficking of released DOX, resulting in elevated cytotoxicity in CathB-overexpressing breast cancer cells. The ARPI-derived bioactive peptides exhibit synergistic anticancer effect with DOX by regulating pro- and antiapoptotic-relevant proteins ( p53, Bax, Bcl-2, pro-caspase-3) at mitochondria. In an orthotopic breast tumor model of CathB-overexpressing breast cancer, DOX-ARPI/CS NPs remarkably inhibit tumor growth, enhance tumor cell apoptosis and prolong host survival without eliciting any systemic toxicity. These results suggest that exploitation of multifunctional biomaterials to specifically produce anticancer agents inside cancer cells and trigger drug release to the subcellular target sites is a promising strategy for designing effective synergistic nanomedicines with minimal off-target toxicity.