| ²é¿´: 662 | »Ø¸´: 1 | |||
| ±¾Ìû²úÉú 1 ¸ö ·ÒëEPI £¬µã»÷ÕâÀï½øÐв鿴 | |||
bxzc123½ð³æ (ÕýʽдÊÖ)
|
[ÇóÖú]
Çó·Ò룬лл£¬Õ¾ÄÚÐÅÇó´óÉñ°ïæ
|
||
|
A novel glyA gene from the marine bacterium Alcanivorax sp. was cloned and expressed in Escherichia coli BL21(DE3). The recombinant glyA encodes a polypeptide of 418 amino acids, which was designated as AdSHMT that shows the highest identity (70%) with a SHMT from Shewanella algae. The purified enzyme showed a single band at about 45 kDa by SDS-PAGE analysis. It was found that AdSHMT exhibited the maximal activity at 50 ◦ C and pH 7.0. The K m , V max , and K cat values of AdSHMT against dl-threo-3-phenylserine were calculated to be 0.097 mol/L, 3.255 ?mol/min/mg and 2.451/s, respectively. More importantly, RP-HPLC detection showed that the AdSHMT achieved an 88.37% molecular conversion rate in catalyzing glycine to l-serine, with the final concentration of l-serine being 353.15 mM in the reaction at 35 ◦ C and 22nd hour when the initial concentration of the substrate (glycine) was 0.399 M. The molecular conversion rate of the AdSHMT from the Alcanivorax sp. was 1.26-fold that of the EcSHMT from the E. coli,which is currently applied in industrial production. Therefore, AdSHMT has the potential for industrial applications due to its high enzymatic conversion rate. |
» ²ÂÄãϲ»¶
302Çóµ÷¼ÁÒ»Ö¾Ô¸»ªÖÐʦ·¶´óѧ
ÒѾÓÐ4È˻ظ´
288Çóµ÷¼Á Ò»Ö¾Ô¸¹þ¹¤´ó ²ÄÁÏÓ뻯¹¤
ÒѾÓÐ32È˻ظ´
Ò»Ö¾Ô¸Ö£´ó²ÄÁϹ¤³Ì290Çóµ÷¼Á
ÒѾÓÐ20È˻ظ´
»·¾³¹¤³Ì297·ÖÇóµ÷¼ÁÒ»Ö¾Ô¸º¼¸ßÔº
ÒѾÓÐ12È˻ظ´
366Çóµ÷¼ÁÒ»Ö¾Ô¸¶«±±´óѧ
ÒѾÓÐ7È˻ظ´
²ÄÁϵ÷¼Á
ÒѾÓÐ6È˻ظ´
Ò»Ö¾Ô¸±±¾©¿Æ¼¼´óѧ085601²ÄÁϹ¤³ÌÓ¢Ò»Êý¶þ³õÊÔ×Ü·Ö335Çóµ÷¼Á
ÒѾÓÐ9È˻ظ´
²ÄÁϵ÷¼Á
ÒѾÓÐ5È˻ظ´
²ÄÁÏÓ뻯¹¤306·ÖÕÒµ÷¼Á
ÒѾÓÐ5È˻ظ´
301Çóµ÷¼Á
ÒѾÓÐ5È˻ظ´
UTKProgress
ľ³æ (СÓÐÃûÆø)
- ·ÒëEPI: 11
- Ó¦Öú: 7 (Ó×¶ùÔ°)
- ½ð±Ò: 3605.5
- É¢½ð: 7
- ºì»¨: 3
- Ìû×Ó: 280
- ÔÚÏß: 236.4Сʱ
- ³æºÅ: 3500502
- ×¢²á: 2014-10-27
- ÐÔ±ð: GG
- רҵ: ¸ß·Ö×Ӻϳɻ¯Ñ§
¡¾´ð°¸¡¿Ó¦Öú»ØÌû
¡ï ¡ï ¡ï ¡ï ¡ï ¡ï ¡ï ¡ï ¡ï ¡ï ¡ï ¡ï ¡ï ¡ï ¡ï ¡ï ¡ï ¡ï ¡ï ¡ï
bxzc123: ½ð±Ò+20, ·ÒëEPI+1, ¡ï¡ï¡ïºÜÓаïÖú 2014-11-25 22:11:57
bxzc123: ½ð±Ò+20, ·ÒëEPI+1, ¡ï¡ï¡ïºÜÓаïÖú 2014-11-25 22:11:57
| Ò»ÖÖÀ´×Ôº£Ñóϸ¾úAlcanivoraxµÄÐÂÐ͵ÄglyA»ùÒò±»¿Ë¡£¬²¢Ôڴ󳦸˾úBL21£¨DE3£©Éϱí´ï¡£ÖØ×éµÄglyA ±àÂëÁËÒ»¸öÓÉ418¸ö°±»ùËá×é³ÉµÄ¶àëÄ£¬±»ÃüÃûΪ AdSHMT£¬ ËüÏÔʾ³öÓëÀ´×ÔShewanellaÔåµÄSHMT×î¸ß70£¥µÄÏàËÆ¡£ÔÚSDS-PAGE·ÖÎöÖд¿»¯µÄøΪ45 kDaµÄµ¥Á´ (¶àëÄ)¡£½á¹û·¢ÏÖ£¬ÔÚ50◦CºÍpH7.0ʱ£¬AdSHMT±íÏÖ³ö×î´óµÄ»îÐÔ¡£AdSHMT against dl-threo-3-phenylserine (ÕâÀïµÄÒ»¸öרҵ֪ʶµã£¬ÎÒ²»¶®¡£Ó¦¸ÃÊÇÒ»Éú»¯·´Ó¦)µÄ Km, Vmax, ºÍ Kcat ÊýÖµ·Ö±ðΪ0097Ħ¶û/Éý£¬3255Ħ¶û/·ÖÖÓ/ºÁ¿ËºÍ2451/Ãë¡£¸üÖØÒªµÄÊÇ·´Ïà¸ßЧҺÏàÉ«Æ×·¨¼ì²â±íÃ÷£¬ÔÚ35 ◦ Cʱ£¨AdSHMT£©´ß»¯glycineΪl-serine·´Ó¦Öе±glycineµÄ³õʼŨ¶ÈΪ0.399 MʱÔÚµÚ22СʱAdSHMTÈ¡µÃÁË88.37%µÄת»¯ÂÊ¡£´ÓAlcanivorax SPµ½AdSHMTµÄ·Ö×Óת»¯ÂÊÊÇĿǰÔÚ¹¤ÒµÉú²úÖÐÓ¦ÓõĴÓE. coliµ½EcSHMTµÄת»¯ÂʵÄ1.26±¶¡£Òò´Ë£¬AdSHMTÒòÆä¸ßµÄøת»¯ÂÊ¿ÉÄÜÔÚ¹¤ÒµÉϵõ½Ó¦Óᣠ|

2Â¥2014-11-25 10:06:15














»Ø¸´´ËÂ¥