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    Unveiling anti-inflammatory peptides from Lion's Mane mushroom (Hericium erinaceus): Preparation, bioactivity assessment, and peptides identification
    (Elsevier B V, 2025-12-01) Silva B; Sringarm K; Potikanond S; Tangjaidee P; Buacheen P; Rachtanapun P; Donlao N; Singh J; Kaur L; Issara U; Kingwascharapong P; Phongthai S
    Lion's Mane mushroom (Hericium erinaceus) is acknowledged worldwide for its substantial contribution of medicinal compounds and nutrients, including protein. The efficient extraction and hydrolysis of proteins are essential for revealing their bioactive properties. This study demonstrates that the optimal pulsed electric field (PEF)-assisted extraction achieved a 42.44 % increase in protein extraction efficiency relative to traditional alkaline extraction (p < 0.05). H. erinaceus protein contained 44.59 % essential amino acids and exhibited 71.33 % in vitro digestibility. Pepsin-trypsin hydrolysis produced the most significant anti-inflammatory activity, resulting in a 36.2 % reduction in nitric oxide and a 31.8 % decrease in interleukin-6 levels (p < 0.05). Subsequent fractionations employing membrane ultrafiltration and size exclusion chromatography effectively purified the peptides, resulting in enhanced anti-inflammatory activity (p < 0.05). This research discovered nine important peptide sequences containing 50–100 % hydrophobic amino acids in Lion's Mane mushroom proteins, which could aid in the synthesis of natural anti-inflammatory peptides.
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    Deodorisation of protein hydrolysate and extraction of proteins from Hoki (Macruronus novaezelandiae) skin : a thesis presented in partial fulfilment of the requirements for the degree of Master of Food Technology at Massey University, Albany, New Zealand
    (Massey University, 2018) Sharim, Nur Syazwana
    The present study had two main objectives. The first objective was to identify a suitable deodorisation treatment for pre-prepared Hoki skin protein hydrolysate (HSPH) and the second objective was to investigated suitable pre-treatment and extraction processes for collagen and gelatine from Hoki (Macruronus novaezelandiae) skin which resulted in low odour extracts. The off-odour in HSPH post-deodorisation treatments and in the Hoki collagen and gelatine post-extraction processes were assessed by determining the total volatile base nitrogen (TVB-N) and trimethylamine (TMA) concentrations. The sensory technique of flash profiling was employed to determine the odour attributes in all HSPH, gelatine and collagen samples after treatment and extraction processes. In addition to the respective off-odour assessments, the extracted collagen and gelatine were evaluated in terms of total protein content (g protein/100g dry sample), moisture content (w/w%), and yield (w/w% of dry sample). Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was conducted to determine the molecular weight (kDa) of extracted collagen and gelatine. Amino acid profile analysis was performed to identify the extracted samples. In the first part of this study, dried green tea leaves (GT), powdered tea polyphenol (TP) and dried olive leaves (OL) investigated for deodorisation of HSPH. Using an orthogonal design, three factors (concentration, temperature and time) and three corresponding levels were used in the design. The two most suitable deodorisation treatments for pre-prepared HSPH were 1) deodorant: powdered tea polyphenol; concentration: 0.04 g/ml hydrolysate; temperature: 50˚C; time: 20 min, and 2) deodorant: powdered tea polyphenol; concentration: 0.04 g/ml hydrolysate; temperature: 80˚C; time: 60 min. For a more economical solution, GT was determined to be a possible alternative deodorant to TP by manipulating the total phenolic content prior to deodorisation. For a secondary deodorisation treatment, preliminary results on strong acid hydrogen form ion exchange resin (Dowex G-26) reduced the TMA concentration in partially deodorised HSPH sample significantly (p-value<0.05) from 3.4±0.1 deodorisation to 0.8±0.1 mg of nitrogen/100g wet sample. In the second part of this study, Hoki skins were pre-treated using 0.2 M NaOH solution (1:6 w/v) for 60 min at 18±2oC and then extraction with distilled water (1:10 w/v) for 60 minutes at 50±2˚C. This treatment produced gelatine product with the highest protein content (41.3±0.9 g of protein/100g dry sample) and reduced off-odour based on TMA content (0.9±0.1 mg of nitrogen/ 100 g wet sample). However, a lower gelatine yield recovery of 61.0±1.7 % was determined in this gelatine sample. SDS-PAGE and amino acid profile tests concluded that all pre-treatment and extraction processes successfully extracted gelatine samples as the final product. In contrast, collagen samples were not confirmed as pure collagen in this study. The current findings for both objectives of this study has shown that pre-treating the raw material using acid or alkali prior to subsequent processes is more efficient in reducing the off-odour in the final products rather than employing deodorisation processes as a subsequent countermeasure after hydrolysis and extraction.
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    Development of a lactic casein based savoury flavour product : a thesis presented in partial fulfilment of the requirement for the degree of Master in Food Technology at Massey University, Manawatū, New Zealand
    (Massey University, 2016) Chen, Ting
    The aim of this project was to develop a casein-based hydrolysate formulation that has higher savoury flavour and is more cost effective than an existing commercial savoury hydrolysate. From the literature review, bovine casein protein has the most savoury flavour potential of all proteins due to its high glutamic acid and glutamine content. The symbol of savoury flavour is cheese which is made from casein protein in the western world. The main reaction resulting in cheese savoury flavour development is proteolysis where casein protein breaks down to peptides by protease and free amino acids by peptidases. Two different systems were designed to be based on those reactions in order to generate maximum free glutamic acid during the experiments. The enzyme substrate was a 10% lactic sodium caseinate slurry, which is the foundation of the experiments. With the first system, an enzyme preparation with protease functions was added first and followed by an enzyme preparation with peptidase functions. With the second system only one enzyme preparation with both protease and peptidase activity was added for each trial. From the results, it was found that none of the enzyme combinations from either system were able to achieve the same amount of free glutamic acid as the existing commercial product (31.95 mg/g of protein) within 24 hours. However, multiple options would have had equivalent free glutamic acid if the free glutamine content could be converted to L-glutamic acid using a glutaminase. Flavorzyme 1000L from system one was selected to be the option combining with glutaminase based on its cost, microbiology and chemistry process results. Two different dosages (0.25% and 0.5%) of Glutaminase C100SD were trialled with 2% of Flavorzyme 1000L. From the degree of hydrolysis, free amino acid content, molecular weight profile and residual glutamine results, there were almost no difference between the two trials. The final formulation of Flavorzyme 2% and 0.25% Glutaminase C100SD had 48% more free glutamic acid than the existing commercial control. It also achieved a 33% ingredient cost reduction. Most importantly, the final formulation resulted in a 22.5% final ingredient cost reduction per kilogram based on the same commercial cost model as the control. An informal sensory panel indicated that the new savoury hydrolysate was more savoury and less bitter than the existing commercial control.
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    Beef hydrolysis by Zyactinase™ enzymes : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy at Massey University, Auckland, New Zealand.
    (Massey University, 2016) Ahmad, Noriza Binti
    Protein hydrolysis is the term that applies to all possible ways of splitting proteins to produce products with lower molecular weight. There is a continuous search for novel products derived from waste materials. In the developed nations considerable amount of meat off-cuts are discarded each year. Utilizing these leftovers by developing new technology for protein recovery and modification and production of a broad spectrum of food ingredients greatly enhances its final value. The aim of this research was to partially hydrolyse beef meat protein with a commercial kiwifruit product called ZyactinaseTM, which is essentially freeze-dried kiwifruit to determine the effect of various processing conditions that influence the extent of beef meat hydrolysis. Secondly to determine the peptide and amino acid profile of the beef meat sample after hydrolysis. Thirdly to determine the relative reaction of ZyactinaseTM on various beef meat protein fractions. This study also aimed to evaluate the rate and the extent of partial enzymic hydrolysis of lean beef using ZyactinaseTM enzymes in order to obtain a better understanding of protein hydrolysis reaction. Lean beef minced was partially hydrolysed using the Zyactinase enzymes for different processing times (up to 360 minutes), temperatures (27°C to 70°C) and varying enzyme concentrations. No pH adjustment on the raw material was carried out except for pH studies. The hydrolysates were collected and analysed for total nitrogen content and degree of hydrolysis. The method used to characterize the extent of protein hydrolysis was SN-TCA index (fraction of nitrogen soluble in trichloroacetic acid) also called non-protein nitrogen NPN. Peptide and amino acid in protein hydrolysates were analysed by HPLC and different protein fractions in the hydrolysates were characterised by SDS-PAGE. The relationship between the reaction temperature, enzyme concentration and processing time to the total nitrogen and NPN were determined. The total nitrogen content remained relatively constant throughout the hydrolysis process. In addition, the NPN content increased as the temperature, processing time and enzyme concentration increased. The optimum pH range for the enzyme’s activity was 4 – 5.6 and optimum temperature was 60°C. Furthermore, most of the higher molecular weight protein bands on SDS- PAGE disappeared after hydrolysis and lower molecular weight protein bands increased in intensity. Zyactinase was also found to digest protein in the myobrilla and sarcoplasmic meat fractions at similar rates as whole beef meat. The results provide basic understanding of the kiwifruit enzymes action toward protein that may lead to improved methods for recovering meat protein or developing new food materials.
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    The effect of enzymatic hydrolysis of a dietary protein on the excretion of urinary nitrogen metabolites : a thesis submitted in partial fulfilment of the requirements for the degree of Master of Science in Nutritional Science at Massey University, Palmerston North, New Zealand
    (Massey University, 2000) Quintino Cintora, Maria Eugenia.
    Hydrolysed milk proteins are used for many purposes in human nutrition. Although it is assumed that the nutritive value of a protein hydrolysate is the same, or even superior to the corresponding intact protein, there is limited research available to support this assumption. The aim of this study was to compare amino acid utilisation and the pattern of excretion in the urine of the nitrogenous metabolites (urea, ammonia and creatinine) as an immediate response to the ingestion of a meal containing an intact protein or its enzymatic hydrolysate. This involved a novel technique, 'acute urine collection' (AUC), in which urine was drained from the bladder at short time periods (30 min to 2 hr) through a catheter. The performance and nitrogen balance results indicated that the two sources of amino acid were equally effective in supporting nitrogen retention and growth of the pigs. Nevertheless, the pattern of excretion of the metabolites of nitrogen digestion suggested important differences in the metabolism of the pigs on the two diets. Both groups of pig excreted creatinine nitrogen, at constant and comparable rates over the sampling period indicating similar rates of catabolism in the muscle. The total excretion of nitrogen by AUC by the two groups was similar but the pattern of excretion over the day differed which indicated a difference in the metabolism of the amino acids in the diets. This may have been in part due to a more rapid absorption of amino acids from the hydrolysed diet and in part due to a higher rate of glutamine and asparagine breakdown in the gut of pigs fed the hydrolysate. Excretion of nitrogen as urea and ammonia was similar for the two groups but there were differences between the groups in the pattern of excretion of these metabolites. In addition, the excretion of ammonia was significantly lower (P <0.0001) in the pigs fed the hydrolysate. This was due to a higher content of fixed cations in the diet containing the hydrolysate that led to a compensatory reduction in ammonia excretion. There was a proportional increase in the excretion of urea in the pigs on the hydrolysed diet as a result of the reduction in ammonia excretion but the differences were small relative to the total urea excretion and not significant. AUC not only gives comparable information to the nitrogen balance if it is carried out over a 24 hr period but it also provides detailed information about the protein utilisation during the immediate postprandial period. In particular, AUC can indicate differences and/or similarities in protein absorption by allowing the observation of the pattern of production of urea directly related to the catabolism of dietary amino acids. In addition, it may be possible to use this technique to estimate the optimum time between meals.