Figure 3-20. SDS-PAGE analysis of proteins in DMW20. The lanes are: Marker, Protein Molecular Weight Marker (Broad); Day 0 W, whole sample of DMW20 before fermentation; Day 16 S, supernatant of DMW20 after fermentation; Day 16 W, whole sample of DMW20 after fermentation.
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Figure 3-1に示したように、発酵後のサンプルにおいて、タンパク質の沈殿が見られた。
Figure 3-20より、沈殿したタンパク質は、主にラクトフェリン、ラクトペルオキシダー
ゼ、免疫グロブリン、カゼインであることが示唆された。また、発酵後の上清のレーン において、バンドが薄くなっているので、一部のウシ血清アルブミン、β-ラクトグロブ リン、およびα-ラクトアルブミンも沈殿したと推測できる。
Yadav氏らの総説によると、ホエイタンパク質のそれぞれの等電点は、ウシ血清アルブ
ミンでpH 5.1付近、β-ラクトグロブリンでpH 5.4付近、α-ラクトアルブミンでpH 4.5
付近、ラクトフェリンとラクトペルオキシダーゼでpH 9.5付近である (Yadav et al., 2015)。
上述した「至適初期pHの決定」における結果および考察では、タンパク質の沈殿には pHだけでなく、エタノールの作用もしくは両方の相互作用が関わっていると考察した。
しかし、ホエイタンパク質の種類によって等電点が異なり、pH の低下によって沈殿す ると推測した β-ラクトグロブリンおよび α-ラクトアルブミンが上清に多く残っている ことから、発酵によって沈殿したタンパク質と、Figure 3-12に示したpHの低下によっ て沈殿したタンパク質とが、異なる可能性が示唆された。また、沈殿したタンパク質や 残存したタンパク質によって、官能特性や香気成分量が異なる可能性がある。ホエイタ ンパク質の安定性に最も影響のある要因は定かではないため、より詳細な研究が必要で ある。
個々のホエイプロテインの重要な機能性と栄養的役割が、Yadav氏らの総説にまとめら れている (Yadav et al., 2015)。ラクトフェリンは抗菌、抗ウイルス、抗かび、および免疫 調整作用、ラクトペルオキシダーゼは抗ウイルスおよび抗炎症作用、免疫グロブリンは 免疫調整および成長発達作用、ウシ血清アルブミンは必須アミノ酸源、β-ラクトグロブ リンは必須アミノ酸源、分枝鎖アミノ酸源、および輸送タンパク質、α-ラクトアルブミ ンは必須アミノ酸源、分枝鎖アミノ酸源、および抗がん作用としての機能を持つ。本研 究で開発した発酵飲料においても、ウシ血清アルブミン、β-ラクトグロブリン、および α-ラクトアルブミンが含まれているため、必須アミノ酸や分枝鎖アミノ酸の栄養源とな りえるので、筋肉の健康に関する機能性が期待できる。
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4.
複合型官能評価と発酵試験とを用いて、最適な原料条件 (原料ホエイと基質濃度)、およ び発酵条件 (至適発酵温度と至適初期pH)、ならびに開発した発酵飲料の保存期間を評 価した。その結果、ホエイ由来発酵飲料の開発における最適な発酵条件は、ホエイ原料 は脱塩ホエイ、基質濃度は20% w/v、発酵温度は30℃、初期pHはpH 7という条件で あると明らかになった。また、本研究で開発したホエイ由来発酵飲料の保存期間は、最 大4週間であるとわかった。本研究において提唱した複合型官能評価は、単一の官能評 価と比べて、最適条件の選択に有用であることが示された。複数の官能評価により、様々 な観点からサンプルを評価でき、嗜好性と強度の相関を明らかにし、判断の根拠を増や すことができた。
最終産物に関して、アミノ酸および香気成分の定量、抗酸化能の評価、残存タンパク質 の確認をおこなった。結果としては、先行研究で開発された、比較的似た条件で作られ た飲料と遜色のないアミノ酸量および香気成分量、ならびに抗酸化能であることがわか った。残存タンパク質として、必須アミノ酸や分枝鎖アミノ酸の栄養源となるタンパク 質が飲料中に含まれていたため、筋肉の健康などのホエイタンパク質に由来する機能の 一部が期待できる。
本研究における、脱塩ホエイを用いた発酵飲料の開発は、添加剤や蒸留などの追加のア プローチが不要であり、純粋培養したGRAS酵母によって単一の基質を発酵できる。そ のため、簡単、便利かつ経済的に、工業利用されていないホエイの割合を減らし、新た な需要や価値を生み出せる可能性がある。
今後は、トレーニングした評価パネルによる官能評価の実施、HPLC-MS やGC-MS に よる詳細な成分分析、さらにそれらを掛け合わせたデータを用いた主成分分析などが必 要である。また、炭酸の溶解、熟成、提供温度などのアプローチは未検討であるため、
発酵飲料の嗜好性をさらに高められる余地がある。
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