Round scad protein-based film: Storage stability and its effectiveness for shelf-life extension of dried fish powder

Storage stability of different round scad (Decapterus maruadsi) protein-based films including (1) control film without palm oil and chitosan, (2) film with 25% palm oil (glycerol substitution) and (3) film with 25% palm oil and 40% chitosan (protein substitution) was investigated. During storage (54% relative humidity, 28-30 °C), tensile strength (TS) of the films without and with 25% palm oil or 25% palm oil in combination with 40% chitosan increased continuously, while elongation at break (EAB) decreased markedly when storage time increased up to 8 weeks (p < 0.05). No changes in water barrier properties were observed throughout the storage time up to 4 weeks (p > 0.05). Film solubility slightly increased but protein solubility decreased continuously during storage (p < 0.05). All films became darker and more yellowish as storage time increased. When round scad muscle protein-based films were used to cover dried fish powder, the samples covered with the film containing 25% palm oil and 40% chitosan showed lower thiobarbituric acid reactive substances (TBARS) and yellowness than other samples during the extended storage up to 21 days (p < 0.05). No differences in moisture content of all samples covered with different films were noticeable (p > 0.05), except those covered with HDPE, which had the lowest moisture content. Thus, round scad protein-based film incorporated with palm oil and chitosan could be a promising packaging material to prevent lipid oxidation in oil enriched foods.     

Properties of a protein-based film from round scad (Decapterus maruadsi) as affected by muscle types and washing

The effects of muscle types and washing on the properties of a protein-based film from round scad (Decapterus maruadsi) mince were investigated. Washing resulted in an increase in the protein content with a coincidental decrease in the fat content of mince, especially from whole muscle and dark muscle. Among all types of muscle (ordinary, dark and whole muscle), the ordinary muscle rendered the film with the highest tensile strength (TS) (p < 0.05). TS of films from washed mince was greater than that of films prepared from unwashed mince for the same type of muscle used (p < 0.05). Nevertheless, the water vapour permeability (WVP) of films from unwashed mince was higher than that of films prepared from washed mince (p < 0.05). Films from washed mince had higher solubility but lower protein solubility than those from unwashed mince (p < 0.05). Regardless of washing, films from ordinary muscle showed the highest L^∗-value (p < 0.05). However, films prepared from dark muscle were more yellowish than those prepared from other muscles, as evidenced by the greater b^∗-value. Films from round scad mince and washed mince had excellent barrier properties to UV light at the wavelength of 200–280 nm. Generally, films from round scad mince had a lower preventive effect on visible light transmission than had those from unwashed mince. Among films from all muscles, those prepared from dark muscle exhibited the highest barrier to visible light transmission (p < 0.05). Therefore, the properties of films from round scad meat were governed by muscle type as well as by washing.     

Properties of biodegradable blend films based on fish myofibrillar protein and polyvinyl alcohol as influenced by blend composition and pH level

Effects of the ratios of fish myofibrillar protein (FMP) from bigeye snapper (Priacanthus tayenus) to polyvinyl alcohol (PVA) (FMP:PVA; 10:0, 8:2, 6:4, 5:5, 4:6, 2:8, 0:10) and pH levels (3 and 11) on the properties of resulting films were investigated. Both tensile strength (TS) and elongation at break (EAB) of films increased with increasing PVA content (p < 0.05). When PVA was incorporated up to 40%, films prepared at pH 11 had the higher TS than did those prepared at pH 3 (p < 0.05). However, as PVA content was greater than 40%, films prepared at pH 3 exhibited the higher TS than did those prepared at pH 11 (p < 0.05). Water vapor permeability (WVP) of the films prepared at pH 3 increased when PVA content increased up to 40% and decreased with further increases in PVA content (p < 0.05). PVA films had the higher TS, EAB and WVP than did FMP films and FMP/PVA blend films prepared at both pHs. Films exhibited the increased L∗ and a∗ values but decreased b∗ value with increasing PVA content at both pHs. Films prepared at pH 11 showed higher b∗ value than did those prepared at pH 3 when PVA content was greater than 40% (p < 0.05). FMP/PVA blend films exhibited the negligible transmission to the UV light. At pH 3, light transmission of the films increased as PVA content increased (p < 0.05). At all FMP/PVA ratios, films prepared at pH 11 were less transparent than those prepared at pH 3 (p < 0.05). Therefore, blend composition and pH level influenced the properties of FMP/PVA blend films.     

Properties of protein-based film from round scad (Decapterus maruadsi) muscle as influenced by fish quality

The properties of film prepared from round scad (Decapterus maruadsi) stored in ice for different times were investigated. Degradation of myosin heavy chain (MHC) was more pronounced with the coincidental increase in total volatile base and trimethylamine contents as the storage time increased (P<0.05). Regardless of storage time, no changes in tensile strength (TS) and elongation at break (EAB) of resulting films prepared from unwashed mince were observed (P>0.05). For the films prepared from washed mince, TS decreased, whereas EAB increased when the storage time of fish increased (P<0.05). However, films prepared from washed mince showed the greater mechanical properties with the lower film solubility and protein solubility than did those from mince (P<0.05). Generally, films prepared from fish stored in ice for a longer time became less transparent, darker and more yellowish. The electrophoretic study revealed that similar protein patterns were observed between films, irrespective of storage time of fish and washing. Therefore, the quality of fish did not show the marked impact on the mechanical property of the resulting films, while washing likely affected the film forming ability.     

Effects of plasticizers on the properties of edible films from skin gelatin of bigeye snapper and brownstripe red snapper

The effects of type and concentration of plasticizers on the mechanical properties (tensile strength, TS and elongation at break, EAB), water vapor permeability, light transmission, transparency and color of fish skin gelatin edible films from bigeye snapper (Priacanthus marcracanthus) and brownstripe red snapper (Lutjanus vitta) were investigated. At the same plasticizer concentration, fish skin gelatin films from both species plasticized with glycerol (Gly) showed the greatest EAB (P<0.05), whereas ethylene glycol (EG) plasticized films showed the highest TS (P<0.05). Films prepared from brownstripe red snapper skin gelatin exhibited slightly greater TS than those of bigeye snapper skin gelatin (P<0.05) when Gly and sorbitol (Sor) were used. EG, polyethylene glycol 200 (PEG 200) and polyethylene glycol 400 (PEG 400) affected the mechanical properties of both films differently. Films generally became more transparent and EAB, water vapor permeability (WVP), as well as light transmission of films increased, but TS and yellowness decreased with increasing plasticizer concentrations.     

Effects of partial hydrolysis and plasticizer content on the properties of film from cuttlefish (Sepia pharaonis) skin gelatin

Properties of film from cuttlefish (Sepia pharaonis) ventral skin gelatin with different degree of hydrolysis (DH: 0.40, 0.80 and 1.20%) added with glycerol as plasticizer at various levels (10, 15 and 20%, based on protein) were investigated. Films prepared from gelatin with all DH had the lower tensile strength (TS) and elongation at break (EAB) but higher water vapor permeability (WVP), compared with the control film (without hydrolysis) (p < 0.05). At the same glycerol content, both TS and EAB decreased, while WVP increased (p < 0.05) with increasing %DH. At the same DH, TS generally decreased as glycerol content increased (p < 0.05), however glycerol content had no effect on EAB when gelatins with 0.80 and 1.20% DH were used (p > 0.05). DH and glycerol content had no marked impact on color and the difference in color (ΔE^∗) of resulting films. Electrophoretic study revealed that degradation of gelatin and their corresponding films was more pronounced with increased %DH, resulting in the lower mechanical properties of films. Based on FTIR spectra, with the increasing %DH as well as glycerol content, higher amplitudes for amide-A and amide-B peaks were observed, compared with film from gelatin without hydrolysis (control film) due to the increased –NH₂ group caused by hydrolysis and the lower interaction of –NH₂ group in the presence of higher glycerol. Thermo-gravimetric analysis indicated that film prepared from gelatin with 1.20% DH exhibited the higher heat susceptibility and weight loss in the temperature range of 50–600 °C, compared with control film. Thus, both chain length of gelatin and glycerol content directly affected the properties of cuttlefish skin gelatin films.     

Properties and antioxidant activity of fish skin gelatin film incorporated with citrus essential oils

Properties of protein-based film from fish skin gelatin incorporated with different citrus essential oils, including bergamot, kaffir lime, lemon and lime (50% based on protein) in the presence of 20% and 30% glycerol were investigated. Films containing 20% glycerol had higher tensile strength (TS) but lower elongation at break (EAB), compared with those prepared with 30% glycerol, regardless of essential oils incorporated (p < 0.05). Films incorporated with essential oils, especially from lime, at both glycerol levels showed the lower TS but higher EAB than the control films (without incorporated essential oil) (p < 0.05). Water vapour permeability (WVP) of films containing essential oils was lower than that of control films for both glycerol levels (p < 0.05). Films with essential oils had varying ΔE^* (total colour difference), where the highest value was observed in that added with bergamot essential oil (p < 0.05). Higher glycerol content increased EAB and WVP but decreased TS of films. Fourier transforms infrared (FTIR) spectra indicated that films added with essential oils exhibited higher hydrophobicity with higher amplitude at wavenumber of 2874–2926 cm⁻¹ and 1731–1742 cm⁻¹ than control film. Film incorporated with essential oils exhibited slightly lower thermal degradation resistance, compared to the control film. Varying effect of essential oil on thermal degradation temperature and weight loss was noticeable, but all films prepared using 20% glycerol had higher thermal degradation temperature with lower weight loss, compared with those containing 30% glycerol. Films added with all types of essential oils had rough cross-section, compared with control films, irrespective of glycerol levels. However, smooth surface was observed in all film samples. Film incorporated with lemon essential oil showed the highest ABTS radical scavenging activity and ferric reducing antioxidant power (FRAP) (p < 0.05), while the other films had lower activity. Thus, the incorporation of different essential oils and glycerol levels directly affected the properties of gelatin-based film from fish skin.     

Fatty acids and their sucrose esters affect the properties of fish skin gelatin-based film

The effects of fatty acids (FA) [palmitic acid (PA) and stearic acid (SA)] and their sucrose esters (FASE) on the mechanical properties, water vapor permeability (WVP), light transmission, and color of films from bigeye snapper and brownstripe red snapper skin gelatins were investigated. Tensile strength (TS) of films generally decreased with the addition of FA (p<0.05), while gradually increased with increasing FASE amount (p<0.05). WVP of films generally decreased with increasing amount of FA or FASE (p<0.05). However, films containing FASE exhibited the superior WVP barrier property to those added with FA. Marked increase in elongation at break (EAB) was observed when either FA or FASE at a level of 25% substitution was incorporated. Light transmission of films in both UV (200–280 nm) and visible ranges (350–800 nm) decreased with increasing FA amount. Films added with FASE were generally more transparent than those with FA. Chain length of FA or FASE affected the properties of films differently, depending upon gelatin sources. Therefore, the properties of fish skin gelatin-based films, especially water vapor barrier, could be improved by the addition of FA or FASE.     

Starch-gelatin edible films: Water vapor permeability and mechanical properties as affected by plasticizers

Physical and mechanical properties of edible films based on blends of sago starch and fish gelatin plasticized with glycerol or sorbitol (25%, w/w) were investigated. Film forming solutions of different ratios of sago starch to fish gelatin (1:0, 2:1, 3:1, 4:1, and 5:1) were used and cast at room temperature. Amylose content of sago starch was between 32 and 34% and the protein content of the fish gelatin was found to be 81.3%. The findings of this study showed that the addition of fish gelatin in starch solutions has a significant effect (p < 0.05), resulting in films with lower tensile strength (TS) and higher water vapor permeability (WVP). On the other hand, increasing protein content (from 10.9% to 21.6%) in film samples plasticized with sorbitol showed significantly lower (p < 0.05) TS but no trend was observed in % elongation-at-break (EAB) and no differences in WVP. However, TS decreased with higher protein content in the samples when either plasticizers were used in general, but no significance differences was observed among the samples (p < 0.05) with glycerol with exception to film with high protein content (21.6%) only and no trend was observed in % EAB among samples as well. Significant difference (p < 0.05) was observed in TS and viscosity between different formulations with sorbitol. The morphology study of the sago starch/fish gelatin films showed smoother surfaces with decreasing protein in the samples with either plasticizer. DSC scans showed that plasticizers and protein content incorporated with sago starch films reduced the glass transition temperature (T_g) and melting temperature (T_m) and the melting enthalpy (ΔH_m). In this study, observation of a single T_g is an indication of the compatibility of the sago starch and fish gelatin polymers to form films at the concentration levels used.     

Physico-chemical properties,morphology and antioxidant activity of film from fish skin gelatin incorporated with root essential oils

The influences of three root essential oils (ginger, turmeric and plai) at different levels (25%, 50% and 100%, based on protein content) on properties and antioxidative activity of fish skin gelatin-based film were investigated. Films incorporated with all essential oils showed the lower tensile strength (TS) but higher elongation at break (EAB) with increasing amount of essential oils, compared with the control film (without oil incorporated), regardless of types of essential oil (p < 0.05). Water vapor permeability (WVP) of films containing essential oils decreased as the amount of essential oils increased (p < 0.05). Decreases in L^*-value and increases in b^*-, ΔE^*- and transparency value were observed with increasing amount of essential oils incorporated (p < 0.05). FTIR spectra indicated that films added with essential oils, especially from plai root, exhibited higher hydrophobicity than the control film, as evidenced by higher amplitude at wavenunber of 2877–2922 cm⁻¹ and 1732 cm⁻¹. Lower degradation temperature was obtained in films containing essential oils. Microstructural study revealed that bilayer films could be formed when essential oils at level above 50% were incorporated. Film incorporated with plai and turmeric essential oils showed the higher DPPH and ABTS radical scavenging activity, respectively, (p < 0.05), compared with the control film and ginger essential oil added film. Thus, the incorporation of root essential oils directly affected properties of fish skin gelatin-based film, depending on types and levels incorporated.     

Roles of lipid oxidation and pH on properties and yellow discolouration during storage of film from red tilapia (Oreochromis niloticus) muscle protein

Protein-based films prepared from red tilapia (Oreochromis niloticus) washed and unwashed mince solubilised at pH 3 and 11 were prepared and characterised. Tensile strength (TS) of films from washed mince was greater than that of films prepared from unwashed mince for both pH used (P < 0.05). TS of films prepared at pH 3 was higher than that of films prepared at pH 11 for both of washed and unwashed mince (P < 0.05). Film from washed mince with pH 3 showed the highest TS, while that from unwashed mince with pH 11 had the lowest TS with the highest elongation at break (EAB) (P < 0.05). Films from washed mince had the lower value of thiobarbituric acid reactive substances (TBARS) than did those from unwashed counterpart, regardless of pH used. Nevertheless, TBARS was much higher in films prepared at acidic pH, compared with those prepared at alkaline pH. During storage of 20 days at room temperature, films became yellowish as evidenced by the increases in b^∗ and ΔE^∗-values. Films prepared at pH 11 showed the higher b^∗ and ΔE^∗-values than did those prepared at pH 3, especially for those from unwashed mince. However, films prepared from washed mince at pH 3 showed higher b^∗ and ΔE^∗-values than did those prepared at pH 11 (P < 0.05). Films generally had the increase in TS but the decreases in water vapour permeability (WVP), film solubility and protein solubility after 20 days of storage (P < 0.05). Therefore, lipid oxidation more likely played a role in yellow discolouration of fish muscle protein film, mainly by providing the carbonyl groups involved in Maillard reaction, while pH regulated the rate of reaction.     

Effect of some factors and pretreatment on the properties of porcine plasma protein-based films

The properties of porcine plasma protein-based films as influenced by some factors and pretreatment were studied. Both protein concentrations (20 and 30 g L⁻¹) and glycerol contents (50, 60 and 70 g/100 g protein) had the impact on film properties. Film prepared from film-forming solution (FFS) containing protein (30 g L⁻¹) and glycerol (60 g/100 g protein) possessed the highest tensile strength (TS) (2.48 MPa), while that containing protein (30 g L⁻¹) and glycerol (70 g/100 g protein) exhibited the greatest elongation at break (EAB) (18.33%). Protein and glycerol contents affected water vapor permeability (WVP) and transparency of the resulting films. No differences in protein solubility were found among all films (p > 0.05). Pretreatment of FFS by adjusting pH (2-11) and heating at different temperatures (40, 55 and 70 °C) on the properties of the resulting films was investigated. TS and EAB became higher but WVP decreased with decreasing or increasing pH value of FFS. Heat treatment of FFS with pH 3 and 10 had no impact on TS of the resulting film (p > 0.05). On the other hand, EAB and WVP increased with increasing temperature of FFS at both pHs (p < 0.05).     

Structure-modifying alkaline and acidic pH-shifting processes promote film formation of soy proteins

The effect of pH-shifting, a process that induces the molten globule state in proteins, on the film-forming potential of soy protein isolate (SPI) at different temperatures was investigated. Partial unfolding at pH 1.5 or 12, followed by refolding at pH 7.0, was performed to alter the protein structure. Glycerin-plasticised films were prepared from pH-treated SPI at ambient temperature (20 °C), or by heating at 50, 60, 70, or 80 °C (30 min). Tensile strength (TS), elongation at break (EAB), water vapour permeability (WVP), protein solubility (pH 3–7), and non-participating proteins of films were analysed, and the film microstructures were examined. The pH₁₂-treated SPI spontaneously formed a transparent, slightly yellowish film at 20 °C, which had the greatest EAB, while pH_1.5-treated and native SPIs required preheating at 50 and 70 °C, respectively, to form a film. Heating generally decreased solubility and WVP but increased TS. Films formed from both pH₁₂- and pH_1.5-treated SPIs were more elastic (up to 2-fold greater in EAB, P < 0.05) than the film formed from untreated SPI despite slightly reduced TS and WVP. Electrophoresis revealed disulphide bonds between A and B subunits of glycinin being a dominant force in pH₁₂- and pH_1.5-treated SPI films, while noncovalent forces were abundant in untreated SPI films. The pH₁₂-treated SPI film consisted of more interactive protein strands than other SPI films, which seemed to explain its superior elastic properties.     

The role of sarcoplasmic protein in hydrostatic pressure-induced myofibrillar protein denaturation

To observe the role of sarcoplasmic protein (SP) on myofibrillar protein (MP) denaturation under a hydrostatic pressure (HP), MP isolated from bovine muscle was treated with 300 MPa by increasing concentrations of SP (0, 0.8, 1.6, and 3.2 mg/ml) from bovine. SDS-PAGE patterns of soluble proteins in 0.1M NaCl (pH 7.4) indicated that a protein (about 100 kDa) from MP decreased with increasing concentrations of SP and that a 97 kDa protein from SP observed with 0.1 MPa was not observed with 300 MPa. SDS-PAGE patterns of soluble proteins in 0.6 M NaCl (pH 7.4) and Ca-ATPase activity showed that the denaturation of myosin heavy chain (MHC) was accelerated with increasing SP concentrations with the 300 MPa treatment. Thus, the addition of SP enhanced HP-induced denaturation of MHC and of a protein from MP of about 100 kDa.     

Effect of phenolic compounds on the properties of porcine plasma protein-based film

Properties of porcine plasma protein-based film incorporated with tannic acid, caffeic acid and ferulic acid at different concentrations (1–3% (w/w) of protein content) were studied. Film-forming solution (FFS) containing 3% protein (w/v) and 70% glycerol (w/w of protein content) was preheated at 70 °C for 30 min and adjusted pH to 10 followed by the addition of phenols and film casting. Tensile strength (TS) of resulting film increased by 123.3, 194.3 and 19.5% and elongation at break (EAB) increased by 71.1, 86.3 and 10.2%, respectively, compared with the control film, when tannic acid, caffeic acid and ferulic acid at a level of 3% was added. The use of all phenolic compounds slightly increased water vapor permeability (WVP) of resulting films (p < 0.05). The increases in a^∗- and b^∗-values of films were observed as the higher concentrations of tannic acid and caffeic acid were used. This was associated with the lowered transparency of resulting films. FFS containing 3% caffeic acid with prior oxygenation, especially with pH 10, yielded the film with increased TS but lowered EAB (p < 0.05). Oxygenation of FFS was associated with the lower L^∗-value and higher a^∗-value of resulting films. Therefore, phenolic compounds could be used as natural cross-linkers which affected the properties of porcine plasma protein-based film differently.     

Influences of degree of hydrolysis and molecular weight of poly(vinyl alcohol) (PVA) on properties of fish myofibrillar protein/PVA blend films

Biodegradable blend films based on fish myofibrillar protein (FMP) and poly(vinyl alcohol) (PVA) were prepared and characterized. PVA with different degrees of hydrolysis (DH) and molecular weights (MW) had the impact on properties of FMP/PVA (1:1, w/w) blend film. The blend films with higher MW of PVA were more tensile resistant, as indicated by the greater tensile strength (TS) and elongation at break (EAB), while the films with PVA of lower DH were more flexible. The blend film with PVA-BP26 (DH: 86–98% mol; MW: 124,000–130,000 g/mol) exhibited the greatest tensile performance and the lowest water vapor permeability (p < 0.05), compared with other films. SEM and FTIR results revealed that FMP and PVA were compatible and their intermolecular interaction was enhanced, providing the blend film with desirable properties. Therefore, incorporation of PVA with appropriated DH and MW could improve the properties of the FMP-based film.     

Compositions,functional properties and antioxidative activity of protein hydrolysates prepared from round scad (Decapterus maruadsi)

Composition, functional properties and antioxidative activity of a protein hydrolysate prepared from defatted round scad (Decapterus maruadsi) mince, using Flavourzyme, with a degree of hydrolysis (DH) of 60%, were determined. The protein hydrolysate had a high protein content (48.0%) and a high ash content (24.56%). It was brownish yellow in colour (L^∗ = 58.00, a^∗ = 8.38, b^∗ = 28.32). The protein hydrolysate contained a high amount of essential amino acids (48.04%) and had arginine and lysine as the dominant amino acids. Na⁺ was the predominant mineral in the hydrolysate. The protein hydrolysate had an excellent solubility (99%) and possessed interfacial properties, which were governed by their concentrations. The emulsifying activity index of the protein hydrolysate decreased with increasing concentration (p < 0.05). Conversely, the foaming abilities increased as the hydrolysate concentrations increased (p < 0.05). During storage at 25 °C and 4 °C for 6 weeks, the antioxidative activities and the solubility of round scad protein hydrolysate slightly decreased (p < 0.05). Yellowness (b^∗-value) of the protein hydrolysate became more intense as the storage time increased but the rate of increase was more pronounced at 25 °C than at 4 °C.     

Characterization of gelatin films prepared from under-utilized blue shark (Prionace glauca) skin

Gelatin film from blue shark (Prionace glauca) skin was investigated in order to utilize what is one of the most serious marine wastes in Japan. Film properties from shark skin such as tensile strength (TS), elongation at break (EAB) were evaluated. The TS of gelatin film from shark skin was affected by the protein concentration (1, 2 and 3%) of the film-forming solution (FFS). TS of the film from a 2% protein FFS was the highest. EAB and water vapor permeability (WVP) increased with increasing FFS protein concentration. WVP of shark skin gelatin was evidently low as compared to gelatin films from other fish. An increase in the FFS protein concentration decreased transparency at almost all wavelengths. Furthermore, opacity at 280 nm was characteristically high as compared to films from bony fish skin. The addition of glycerol improved flexibility and enhanced the UV barrier property at 280 nm. However, transparency at the visible range and WVP increased with increasing glycerol content.From the above, it was suggested that shark skin gelatin film technology can be applied to pharmaceutical products or rich-fat food due to its excellent water and UV barrier properties.     

Characteristics and antioxidant activity of leaf essential oil–incorporated fish gelatin films as affected by surfactants

Characteristics of film from fish skin gelatin incorporated with four leaf essential oils (lemongrass, basil, citronella and kaffir lime), as influenced by different surfactants (Tween‐20, Tween‐80 and soy lecithin), were investigated. Films incorporated with all essential oils had lower tensile strength with higher elongation at break and thickness, compared with control film (P < 0.05). Films added with Tween‐20 showed higher TS, compared with those containing other surfactants (P < 0.05). Water vapour permeability of films incorporated with all essential oils markedly decreased in comparison with control (P < 0.05). Films generally became darker and more yellowness, when incorporated with essential oils. Film containing basil essential oil had the highest DPPH radical– and ABTS radical–scavenging activities, compared with those added with other essential oils. Higher antioxidative activity was obtained in films containing essential oils when soy lecithin was used as surfactant, probably due to the combined effect of both constituents.     

Potential protein degradation balance and total metabolizable protein supply to dairy cows from heat‐treated faba beans

The effects of pressure toasting (100, 118 and 136 °C for 3, 7, 15 and 30 min) on potential protein nutritional value of faba beans were evaluated with the NRC 2001 dairy model, by determining undegraded (RUP) and degraded rumen protein (RDP), undegraded (RUST) and degraded rumen starch (RDST), truly absorbed undegraded protein (ARUP), microbial protein (MCP_RDP) synthesized in the rumen from rumen‐available protein, truly absorbed rumen synthesized microbial protein (AMCP), truly absorbed rumen endogenous protein (AECP), total metabolizable protein (MP) in the small intestine, and the protein degradation balance (PDB). The treatments increased RUP, RUST, ARUP and MP (p < 0.001), and decreased RDP, RDST, MCP_RDP and PDB (p < 0.001), the effects increasing with increasing temperature and time. The treatments increased (p < 0.001) ARUP without affecting AECP and AMCP, so that the net absorbable total MP in the small intestine was increased. The PDB was reduced (p < 0.001) but never became negative. These results indicated that potential microbial protein synthesis would not be impaired due to sufficient nitrogen in the rumen, but the high positive PDB values with most treatments, except 136 °C for 15 min (PDB 2.0 g kg^?1 DM) indicated that there were large potential losses of nitrogen in the rumen, particularly for the control with a value of 88.9 g kg^?1 dry matter. It is concluded that predicted potential protein degradation balance and total metabolizable protein supply from faba beans were improved by the treatments. Copyright © 2005 Society of Chemical Industry