Efficient Bioproduction of Human Milk Alpha-Lactalbumin in Komagataella phaffii,Journal of Agricultural and Food Chemistry

当前位置： X-MOL 学术 › J. Agric. Food Chem. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Efficient Bioproduction of Human Milk Alpha-Lactalbumin in Komagataella phaffii
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2022-02-11 , DOI: 10.1021/acs.jafc.1c07908
Mengting Deng _{1,

2,

3,

4} , Xueqin Lv _{1,

2,

3,

4} , Long Liu _{1,

2,

3,

4} , Jianghua Li _{1,

2,

3,

4} , Guocheng Du _{1,

2,

3,

4} , Jian Chen _{2,

3,

4,

5,

6} , Yanfeng Liu _{1,

2,

3,

4,

6}

Affiliation

Alpha-lactalbumin (α-LA; the most abundant whey protein in human milk) contributes to infant development, providing bioactive peptides and essential amino acids. Here, Komagataella phaffii (K. phaffii) was selected as the production host. We found that the K. phaffii host X33 was suitable for expressing the target protein, yielding 5.2 mg·L^–1 α-LA. Thereafter, several secretory signal peptides were applied to obtain a higher titer of α-LA. The strain with α-factor secretory signal peptide secreted the highest extracellular titer. Additionally, promoters AOX1, GAP, and GAP(m) were compared and applied. The strain with the promoter AOX1 produced the highest extracellular titer. In addition, coexpressing human protein disulfide isomerase A3 (hPDIA3) increased the titer by 27%. Human α-LA production by the strain X33-pPICZαA-hLALBA-hPDIA3 reached 56.3 mg·L^–1 in a 3 L bioreactor. This is the first report of successful secretory human α-LA expression in K. phaffii and lays foundations for the simulation of human milk for infant formulas and further development of bioengineered milk.

中文翻译：

Komagataella phaffii 母乳 α-乳清蛋白的高效生物生产

α-乳清蛋白（α-LA；人乳中最丰富的乳清蛋白）有助于婴儿发育，提供生物活性肽和必需氨基酸。在这里，Komagataella phaffii ( K. phaffii ) 被选为生产宿主。我们发现K. phaffii宿主X33 适合表达目标蛋白，产生5.2 mg·L ^–1 α-LA。此后，应用几种分泌信号肽以获得更高滴度的α-LA。具有α因子分泌信号肽的菌株分泌的细胞外滴度最高。此外，比较并应用了启动子AOX1、GAP和GAP(m) 。具有启动子AOX1的菌株产生最高的细胞外滴度。此外，共表达人类蛋白质二硫键异构酶 A3 (h PDIA3 ) 使滴度增加了 27%。在 3 L 生物反应器中，菌株 X33-pPICZαA-hLALBA-hPDIA3 产生的人类 α-LA 达到 56.3 mg·L ^{–1 。}这是在法菲克氏酵母中成功分泌人α-LA表达的第一份报告，为婴儿配方奶粉的母乳模拟和生物工程乳的进一步开发奠定了基础。

更新日期：2022-02-11

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南