Study on the functional enhancement of fish collagen by glycation using the Maillard reaction [an abstract of entire text]

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Study on the functional enhancement of fish collagen by glycation using the Maillard reaction [an abstract of entire text]

楊, 博学

北海道大学. 博士(水産科学) 甲第15255号

2023-03-23

http://hdl.handle.net/2115/89922

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theses (doctoral - abstract of entire text)

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主論文の要約
博士の専攻分野の名称：博士（水産科学）

氏名：楊

博学（Yang

Boxue）

学 位 論 文 題 目
Study on the functional enhancement of fish collagen by glycation using the Maillard reaction
（メイラード反応を用いた糖修飾による魚類コラーゲンの機能性増強に関する研究）

The objectives of this study are to investigate the improving effect of the Maillard reaction on
the antioxidant activity and anti-inflammation activity of collagen, to understand the relationship
between the functional improving effect and progress of the Maillard reaction. In order to expand the
application and proceeding valorization of fish collagen, the author focused on the relation between
the enhanced antioxidant activity of collagen with the Maillard reaction proceeding as well as the
mechanism of the increased anti-inflammatory of collagen undergoing the Maillard reaction.
This study consists of six chapters: Chapter I presented the general introduction of this work;
Chapter II and Chapter III described the enhanced antioxidant and anti-inflammatory activities of
tilapia scale collagen by glycation via the Maillard reaction; Chapter IV investigated the improved
antioxidant and anti-inflammatory activities of glycated sturgeon notochord collagen hydrolysates.
Chapter V and Chapter VI included the general discussion and conclusions.
Chapter I presented the background of this study. Concretely, collagen as a crucial protein
resource has been applied in various areas, like the food industry and biomedical area. The utilization
of collagen extracted from fish processing by-products derived increasing attention because it is
consistent with humanity`s interest. Additionally, tilapia scale and sturgeon notochord, two typical
fish processing by-products, are promising materials for type I and II collagen extraction, respectively.
On the other hand, developing and enhancing protein functionality is in the spotlight of the food
science area as it fits consumers` demands. Moreover, the Maillard reaction is a brilliant protein
modification method to enhance its health-beneficial functions, such as antioxidant and antiinflammatory activities.
Chapter II characterized the glycated tilapia scale collagen and investigated the antioxidant
activity of the glycated tilapia scale collagen. Specifically, lyophilized tilapia scale collagen was
mixed with a half weight of alginate oligosaccharide (AO) or glucose (Glc) and incubated at 60 °C
and 35% relative humidity for up to 18 h to produce the Maillard-type glycated collagen (C-AO and
C-Glc, respectively). As glycation progressed, the amount of attached sugar coupled with UV-vis
absorbance at 294 nm and 420 nm increased more rapidly in C-Glc than in C-AO, and the available
lysine decreased rapidly in C-Glc compared with C-AO. The early-to-middle- and late-stage products
of the Maillard reaction were involved in enhanced antioxidant activity of C-AO and C-Glc,
respectively. Additionally, C-AO acquired antioxidant activity without marked available lysine loss.
Moreover, the cytoprotective effect of collagen in hydrogen peroxide (H2O2)-induced cell oxidative
1

damage was enhanced by glycation, achieved by reducing malondialdehyde (MDA) content and
increasing superoxide dismutase (SOD) and catalase activities (CAT).
Chapter III investigated the anti-inflammatory activity of glycated tilapia scale collagen. The
C-AO and C-Glc were prepared as the previous description, and were employed in the
lipopolysaccharides (LPS)-stimulated RAW 264.7 macrophage model to examine the antiinflammatory activity. The results presented the anti-inflammatory activity of collagen in the
reduction of inflammation mediators (TNF-α, IL-6, IL-1β, and NO). Additionally, the antiinflammatory activity of collagen was enhanced with AO glycation for 18 h (C-AO18h), which was
maintained until 24-h glycation. Contrarily, C-Glc shows no enhanced anti-inflammatory activity
along all the glycation periods. The enhanced anti-inflammatory activity of C-AO18h was induced
by depressed the LPS receptor (Tlr4 and Cd14) and Myd88 mRNA expression. Moreover, compared
with row collagen, C-AO18h treatment significantly up-regulated the activity of endogenous
antioxidant enzymes (SOD and CAT) and reduced the MDA level in the LPS-stimulated macrophages.
The enhanced antioxidant activity of C-AO18h in the inflammation stage could contribute to the antiinflammatory activity.
Chapter IV explored the extensiveness of applying the Maillard reaction to enhance the healthbeneficial functions (antioxidant and anti-inflammatory functions) of collagen in varied reaction
conditions. In this chapter, the protein resource was changed from tilapia scale collagen (Chapter II
and Chapter III) to sturgeon notochord collagen-papain-hydrolysates (CP). The glycation condition
was changed from 60 °C with 35% relative humidity (Chapter II and Chapter III) to 80 °C with
75% relative humidity (CP-sugars). In addition, glucuronic acid (GlcUA, uronic acid), a derivative
of Glc, was employed as a modified sugar. The reason for the employment of GlcUA is that the
carboxyl groups contained in GlcUA and AO were found to contribute to enhancing the antiinflammatory action. The results of amino acid analysis illustrated that GlcUA had a higher reactivity
to the reactive amino acids than Glc and AO. The health-beneficial functions of CP-sugars were
evaluated by the ABTS radical scavenging assay and by quantification of inflammatory compound
(TNF-α and NO) production in LPS-stimulated macrophages, respectively. Results indicate that the
antioxidant and anti-inflammatory functions of CP-sugars were increased with the glycation
processes, and AO-glycation enhanced the health-beneficial functions of CP without a marked loss
of reactive amino acids compared with the Glc- and GlcUA-glycations. Moreover, contenting of
carboxyl groups in reducing sugar is not a determining factor but could contribute to health-beneficial
function enhancement during the Maillard reaction.
In conclusion, as described in Chapter V (General discussion) and Chapter VI (Conclusions
and prospective), this study successfully created novel multifunctional fish collagens by glycation
with reducing sugars using the Maillard reaction. Additionally, the health-beneficial functions of fish
collagen were significantly enhanced by AO-glycation. The AO-glycation displayed function
improvements without both excessive losses of lysine and remarkable glycation processing, which
reduced the risk of the Maillard reaction application. Moreover, this study proved the extensiveness
of applying the Maillard reaction to enhance the health-beneficial functions of collagen-related
ingredients in various reaction conditions. The achievements of the present study will contribute to
the expend the utilization of collagen-related ingredients as food and biomaterials and to valorize the
fish processing by-products. ...

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