Page 64 - 《精细化工》2023年第4期

g750g ㇫㏳ࡃጒ FINE CHEMICALS す 40 ࢤ

[8]
͆⅕䚥ϔ䛼 [42] ȡ䭵ᬻϛぶ 䛴⩕ڞ㶕䓫⮱᫦∂喑ᰭ㏵ ȕ-͆⅕䚥⮱ϔ䛼䓫ݝγ 200.81 g/L喑䒙ࡃ⢴ᣒ䓾 100%ȡ

ఫ 7 ࣹ䚣㏔㖁ࣺᏁݣิ ȕ-͆⅕䚥
Fig. 7 Preparation of ȕ-alanine by double enzyme cascade

㶕 8 ࣹ䚣㏔㖁ࣺᏁݣิ ȕ-͆⅕䚥⮱᫦∂ࣷϔ䛼
Table 8 Preparation method and yield of ȕ-alanine by double enzyme cascade
Methods Substrates Enzyme Source of panD Host Plasmid Production/(g/L) Yield/% References
Coexpression Fumaric acid AspA E. coli E. coli pET28a喍+喎 80.4 95.3 [48]
BL21(DE3)
ADC Corynebacterium glutamicum pET28a喍+喎
Coexpression Fumaric acid AspA E. coli E. coli ÿ 118.6 ÿ [42]
BL21(DE3)
ADC Bacillus subtilis ÿ
In vitro Fumaric acid AspA E. coli E. coli pET28a喍+喎 8.02 90.0 [49]
BL21(DE3)
ADC Corynebacterium glutamicum pET24a喍+喎
Coexpression Fumaric acid AspA E. coli K12 substrain MG1655 E. coli pRSFDuet-1 200.81 100.0 [8]
BL21(DE3)
ADC Corynebacterium glutamicum pRSFDuet-1
∕喟Ąÿą㶕⹧᱗ᰶⰥڠ្䖀ȡ
3.3 ̶䚣㏔㖁∂ݣิ ȕ-͆⅕䚥 faecalis喎ȠL-๖ۙ⅕䚥䚣喍Გ⎽λ E. coli喎হ L-๖
ȕ-͆⅕䚥᭜व᜽䒲䚣 A ⮱䛺㺮ݺҀ喑䒲䚣 A ౕ ۙ⅕䚥-Į-㙞㓔䚣喍Გ⎽λ C. glutamicum喎̶䚣֣㖁
ߕ➖ȠḺ➖ࣷᓛ⩌➖⮱А䅏͚౴ࣾᡒ䛺㺮҉⩕ȡѳ ݣิ ȕ-͆⅕䚥喑ᰭ㏵㣤ᓄγ 147.4 g/L ⮱ ȕ-͆⅕䚥ϔ
[9]
᭜喑ᄦ᠌ࢄ㟒 Arabidopsis ധ఍㏱͚᝭ᰶ∕䛷㯸⮪䔈 䛼ȡWANG ぶ ᄳ᳜㡶㟪ႏᱳ㣹喍B. subtilis喎Გ⎽
㵹ܳᲽ喑Ꭳ⇎ᰶࣾ⣝̻๔㗍ᱳ㣹 L-๖ۙ⅕䚥-Į-㙞㓔 ⮱ panD ധ఍হ䊑᠌䅤Ⰴ喍T. castaneum喎Გ⎽⮱ panD
䚣㐀Ჱㆨѩ⮱ह⎽㯸⮪喑ڣЃḺ➖Όह᠌ࢄ㟒ㆨѩ喑 ധ఍ౕ E. coli BW25113 ͚ڞ㶕䓫喑Ჱᐧγ㘪㶕䓫͑
఍ₑ喑ᣕ≸Ḻ➖͚⇎ᰶ panD ധ఍ [50] ȡ䔈̭ₒⵁ⾣ࣾ ⻺̺हχಸ L-๖ۙ⅕䚥-Į-㙞㓔䚣⮱䛺㏱㣹喠䕇䓴ࣺ
⣝喑Ḻ➖͚⮱ ȕ-͆⅕䚥ज⩞ᅬ୔ણ㏼䓴 3 ₒ䚣䭺㼐 Ꮑ᲎Ухࡃ喑λ 500 mL ࣺᏁҀ㈨͚㣤ᓄγ 271.5 g/L
㣤ᓄ喑ѳ䄒䕁ᒱ⮱Ꮒ➖䒙ࡃ⢴䔉ѻλ L-๖ۙ⅕䚥-Į-㙞 ⮱ ȕ-͆⅕䚥ȡᰭऻ喑ᄳ͑⻺̺हχಸ L-๖ۙ⅕䚥-Į-
㓔䚣ࣺᏁ䕁ᒱ喠ࣺ́ᏁݺҀᅬ୔ણ䒰ͧᬯ䉢喑Вₑͧ 㙞㓔䚣⮱㑃⴮ധ఍̻ E. coli Გ⎽⮱ AspA ౕ E. coli
Ꮒ➖ݣิ ȕ-͆⅕䚥᜽᱙䒰倅喑̺ڤิጒ͇Ꮑ⩕Фթȡ BL21(DE3)͚ڞ㶕䓫Ꭳ䒙ࡃჹ侙䚥喑ȕ-͆⅕䚥⮱ᰭ倅
䮑γВᅬ୔ણͧᏂ➖喑♓Ꮃ᝺ぶ [51] В侙Გ䚥ͧ ϔ䛼ͧ 200.3 g/L喑䒙ࡃ⢴倅䓫 90.0%ȡ̶䚣㏔㖁∂
࣌᫆喑ݖ⩕ 侙Გ䚥ᐯᲱ 䚣喍Გ⎽λ Alcaligenes ݣิ ȕ-͆⅕䚥⮱Ⱕڠ᪝ᢛຯ㶕 9 ᝭⹧ȡ

㶕 9 ̶䚣㏔㖁∂ݣิ ȕ-͆⅕䚥ࣷϔ䛼
Table 9 Preparation method and yield of ȕ-alanine by three enzyme cascade
Production Yield
Methods Substrates Enzyme Gene source Host Plasmid References
/(g/L) /%
Coexpression Fumaric AspA E. coli E. coli BW25113 pYB1s 200.3 90.0 [9]
acid ADC Bacillus subtilis
ADC Tribolium castaneum
In vitro Maleic Maleate isomerase Alcaligenes faecalis Alcaligenes faecalis ÿ 147.4 96.0 [51]
acid AspA E. coli E. coli ÿ
ADC Corynebacterium glutamicum Corynebacterium glutamicum ÿ
∕喟Ąÿą㶕⹧᱗ᰶⰥڠ្䖀喠Alcaligenes faecalis ͧ㇗ϔⷞ㣹ȡ

3.4 䚣∂ݣิ ȕ-͆⅕䚥⮱䄰ᣔᱧ⤳ࣷᒞ৺఍㉍ L-๖ۙ⅕䚥-Į-㙞㓔䚣ᕔ㘪ࣷיࡃ䒙ࡃ᲎У͑᫦䲏ȡL-
Ⱋݺ喑䚣∂ݣิ ȕ-͆⅕䚥⮱ᒞ৺఍㉍ͨ㺮䯳ౕ͚ ๖ۙ⅕䚥-Į-㙞㓔䚣ͨ㺮ܳͧ͑ㆨ喑̭⻺ᆋλ͆䚛䚝ധ